Endophytic fungi from mangrove inhibit lung cancer cell growth and angiogenesis in vitro

Corrigendum in: /10.3892/or.2023.8608

  • Authors:
    • Xin Liu
    • Xin Wu
    • Yuefan Ma
    • Wenzhang Zhang
    • Liang Hu
    • Xiaowei Feng
    • Xiangyong Li
    • Xudong Tang
  • View Affiliations

  • Published online on: January 16, 2017     https://doi.org/10.3892/or.2017.5366
  • Pages: 1793-1803
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The secondary metabolites of mangrove-derived endophytic fungi contain multiple substances with novel structures and biological activities. In the present study, three types of mangrove plants, namely Kandelia candel, Rhizophora stylosa and Rhizophoraceae from Zhanjiang region including the leaves, roots and stems were collected, and endophytic fungi were isolated, purified and identified from these mangrove plants. MTT assay was used to observe the effects of the isolated endophytic fungi on the growth of A549 and NCI-H460 lung cancer cells. The effect of the endophytic fungi on lung cancer angiogenesis in vitro induced by the HPV-16 E7 oncoprotein was observed. Our results showed that 28 strains of endophytic fungi were isolated, purified and identified from the three types of mangrove plants. Ten strains of endophytic fungi significantly suppressed the growth of A549 and NCI-H460 cells. The average inhibitory rates in the A549 cells were 64.4, 59.5, 81.9, 43.9, 58.3, 56.2, 48.3, 42.4, 93.0 and 49.7%, respectively. The average inhibitory rates in the NCI-H460 cells were 41.2, 49.3, 82.7, 40.7, 53.9, 52.6, 56.8, 64.3, 91.0 and 45.6%, respectively. Particularly, three strains of endophytic fungi markedly inhibited HPV-16 E7 oncoprotein‑induced lung cancer angiogenesis in vitro. These findings contribute to the further screening of potential chemotherapeutic agents from mangrove-derived endophytic fungi.

Introduction

In 1904, the first strain of endophytic fungus was discovered. Since that date, more and more endophytic fungi have been isolated from different natural plants including mangroves (15). Research has demonstrated that the secondary metabolites from endophytic fungi contain multiple bioactive compounds with novel structures (13). The advantages of endophytic fungi include their short culture cycle, mild conditions for growth, little exposure to environmental pollution and easy industrialization. Thus, these fungi can facilitate the development of traditional pharmaceutical agents. Therefore, the identification of novel natural drugs from endophytic fungi living in different environments and ecosystems offers numerous opportunities.

Mangroves live in high saline and humid environments. Thus, numerous bioactive compounds can be generated under these environmental conditions (4,5). A large number of bioactive compounds have been isolated from endophytic fungi of mangroves, such as, flavonoids, alkaloids, terpenes, quinones, cyclic peptide compounds and fatty acids. Recently, an increased number of novel bioactive compounds have been obtained from mangrove-derived endophytic fungi, such as, novel cyclohexenone, cyclopentenone and xanthone derivatives (6), polyketides (nectriacids A-C and 12-epicitreoisocoumarinol) (7), 3-epiarigsugacin E (8), aspergifuranone and isocoumarin derivatives (9), (R)-3-demethyl purpurester A (9), aromatic butyrolactones (flavipesins A and B) (10) and sulfide diketopiperazine derivatives (penicibrocazines A-E) (11). Multiple bioactive compounds from mangrove-derived endophytic fungi have been demonstrated to exhibit inhibitory activities against acetylcholinesterase (AchE) (8), α-glucosidase (9), bacteria (1012) and viruses (13). Particularly, accumulating evidence indicates that various bioactive compounds from mangrove-derived endophytic fungi display antitumor activities (1318). Furthermore, studies have reported the underlying mechanisms of the antitumor effects of mangrove-derived endophytic fungi (1924). An anthraquinone compound G503, isolated from the secondary metabolites of the mangrove endophytic fungus Nigrospora sp. (no. 2508), was reported to induce apoptosis in gastric cancer SGC7901 cells through the mitochondrial pathway (19). Xyloketal B, a marine compound obtained from the mangrove fungus Xylaria sp. (no. 2508) from the South China Sea, was demonstrated to suppress the proliferation and migration of glioblastoma U251 cells by inhibiting the TRPM7-mediated PI3K/Akt and MEK/ERK signaling pathways (20). The marine anthraquinone derivative SZ-685C, isolated from the secondary metabolites of the mangrove endophytic fungus Halorosellinia sp. (no. 1403), was found to induce apoptosis in primary human non-functioning pituitary adenoma (21), breast cancer (22) and human nasopharyngeal carcinoma (NPC) cells (23), and reverse the adriamycin-resistance in breast cancer cells (24) by the inhibition of the Akt pathway. Taken together, these findings indicate that the bioactive compounds from mangrove-derived endophytic fungi can inhibit cancer cell growth via the induction of apoptosis and the inhibition of the Akt signaling pathway. However, the effect of mangrove-derived endophytic fungi on cancer angiogenesis has not yet been reported.

Angiogenesis, the development of new microvascular networks, is required for cancer invasion and metastasis and plays a key role in controlling the development and progression of a variety of cancers (25). The inhibition of cancer angiogenesis can suppress the development and progression of cancers. Therefore, the screening of angiogenic inhibitors from mangrove-derived endophytic fungi is extremely important for identifying new chemotherapeutic drugs for the prevention and treatment of cancers. There are abundant resources of mangroves in Zhanjiang. Therefore, the present study was to isolate, purify and identify endophytic fungi from Zhanjiang mangroves and explore their effects on the growth and angiogenesis of lung cancer cells.

In the present study, we isolated, purified and identified 28 strains of endophytic fungi from three types of mangrove plants, namely Kandelia candel, Rhizophora stylosa and Rhizophoraceae, and 10 strains of endophytic fungi significantly suppressed the growth of lung cancer cell lines, A549 and NCI-H460. Furthermore, to the best of our knowledge, we demonstrated for the first time, that three strains of endophytic fungi markedly inhibited lung cancer angiogenesis in vitro.

Materials and methods

Reagents

Glucose was purchased from the Tianjin Fuchen Chemical Reagents Factory (Tianjin, China). Potato dextrose agar (PDA) was obtained from Hangzhou Microbial Reagent Co., Ltd. (Hangzhou, China). Tryptone and yeast extract reagents were purchased from Oxoid Ltd. (Basingstoke, Hampshire, UK). Transfection reagent (Lipofectamine™ 2000) was obtained from Invitrogen Corp. (Carlsbad, CA, USA). In vitro angiogenesis assay kit (ECM625) was obtained from Millipore (Temecula, CA, USA). Fungus genomic DNA extraction kit was purchased from Bioer Technology Co., Ltd. (Tokyo, Japan). MTT kit was purchased from Beyotime Biotechnology (Shanghai, China).

Collection of mangrove plants

The healthy leaves, roots and stems of mangroves (Kandelia candel, Rhizophora stylosa and Rhizophoraceae) were collected from Haibin Park and Gaoqiao National Mangrove Nature Reserve (Zhanjiang, Guangdong, China). The collected leaves, roots and stems of the mangroves were washed for 2 h under running tap water and were cut into ~0.5 × 0.5 cm pieces within 72 h after collection. The surface of the fragments was sterilized by sequential immersion in 75% ethanol (C2H5OH) for 45 sec and 5% sodium hypochlorite (NaClO) for different times (leaves for 3 min, roots for 10 min and stems for 5 min), followed by washing four to five times with sterile distilled water.

Isolation, purification and culture of the endophytic fungi

The sterilized fragments of mangroves were dried under sterile conditions. The dried fragments were cultured in plates with PDA medium (potato extract 10.0 g/l, glucose 20.0 g/l, agar 13.0 g/l and chloramphenicol 0.1 g/l) at 28°C, and the growth of the endophytic fungal colonies from the mangrove fragments was monitored every day. The fungal colonies which grew out from the mangrove fragments were isolated and transferred to other plates with PDA medium for purification. The purified endophytic fungal colonies were photographed.

Next, the purified endophytic fungal colonies were fermented at 28°C in glucose peptone yeast (GPY) extract broth (tryptone 2.0 g/l, yeast extract 1.0 g/l, glucose 10.0 g/l, and sea salt 20.0 g/l) in a shaking incubator (160 rpm) at 28°C in the dark. Seven days later, fungus culture media were filtered using nylon nets to separate the mycelia and the culture broth. Mycelia were identified by molecular analysis of the internal transcribed spacer (ITS) of the genomic DNA. The culture broths were sterilized by filtration through a 0.22-µm Millipore filter, and the filtrates were used for MTT and in vitro angiogenesis assays.

Molecular identification of the endophytic fungi

Genomic DNA was extracted from the separated mycelia according to the manufacturer's instructions (Bioer Technology Co., Ltd.). 18S rDNA fragments were amplified by PCR methods with universal primers. PCR primers used were: 5′-TCCGTAGGTGAACCTGCGG-3′ (forward) and 5′-TCCTCCGCTTATTGATATGC-3′ (reverse) (GenBank, NM_006486.2). The primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). The thermocycling conditions were as follows: 94°C for 5 min, followed by 35 cycles at 94°C for 45 sec, 55°C for 45 sec, and 72°C for 60 sec, finally 72°C for 7 min. The PCR products were detected by 1.5% agarose gel electrophoresis and DNA sequencing. The results of agarose gel electrophoresis were photographed. DNA sequences were analyzed by Sangon Biotech Co., Ltd. 18S rDNA fragment sequences of the isolated endophytic fungi were compared with those in the GenBank database using BLAST at the National Center for Biotechnology Information (NCBI; Bethesda, MD, USA), and endophytic fungi were classified by morphologic traits and molecular identification. The phylogenetic trees were constructed by Mega 5.0 software.

Cell culture

Human lung adenocarcinoma cell line A549 and human umbilical vein endothelial cells (HUVECs) were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). Human lung cancer cell line NCI-H460 was obtained from the Chinese Academy of Sciences Cell Bank of Type Culture Collection (Shanghai, China). A549 and NCI-H460 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS). HUVEC cells were grown in DEME media containing 10% FBS. All cells were maintained in a 5% CO2 incubator at 37°C.

Transient transfection

Transient transfection was carried out according to a previously described method (26,27). The plasmid (p-EGFP-N1-HPV-16 E7), constructed by our laboratory, was transiently transfected into A549 and NCI-H460 cells using Lipofectamine™ 2000 according to the manufacturer's instructions, wherein transfection with the empty vector (p-EGFP-N1) served as the negative controls. The cells exposed to transfection reagent alone served as mock transfection controls. The transfection efficiency was evaluated by observing green fluorescence under a fluorescence microscope, and the expression of HPV-16 E7 oncoprotein was confirmed in our previous studies (26,27).

MTT assay

The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was performed to determine the effects of endophytic fungi on the growth of A549 and H460 cells. A cell suspension was added to 96-well plates at the density of 5×104/ml (100 µl/well), and cultured in a CO2 incubator overnight at 37°C. Afterwards, the culture medium was replaced with fresh medium, and 20 µl culture broth of the different endophytic fungi was added into the wells for culture at 37°C for 72 h. Each culture broth of the endophytic fungi was repeated four times. Seventy-two hours later, the supernatant was removed, followed by addition of 20 µl MTT [5 mg/ml in phosphate-buffered saline (PBS)] into each well, and the cells were incubated for an additional 4 h. After removing the supernatant, 150 µl dimethyl sulfoxide (DMSO) was added into each well. The cells were incubated for 10 min at room temperature in order to fully dissolve the formed crystals. Absorbance values at 595 nm were measured using a microplate reader and cell viability was calculated.

In vitro angiogenesis assay

An in vitro angiogenesis assay kit (ECM625) was employed to analyze the formation of capillary tube-like structures according to the manufacturer's instructions. Briefly, HUVECs were seeded at a density of 5×103 cells/well onto the surface of 96-well cell culture plates pre-coated with polymerized ECMatrix™. Subsequently, the conditioned media, derived from HPV-16 E7-transfected A549 or NCI-H460 cells treated with 20 µl culture broth of the different endophytic fungi, were respectively added into different wells. Tubule formation was observed under a phase-contrast microscope, and Scion image software was used to analyze the total tube length in three random view fields/well, and the average value was calculated. The experiment was repeated in triplicate.

Statistical analysis

The experiment was repeated at least three times. One way ANOVA and LSD were employed for statistical analysis using SPSS 19.0. P<0.05 was considered to indicate a statistically significant result.

Results

Results of the isolation, purification and identification of the endophyte fungi

Sixty-two strains of endophytic fungi were isolated from three types of mangrove plants (Kandelia candel, Rhizophora stylosa and Rhizophoraceae) in the Zhanjiang region. The number of endophytic fungus strains isolated from Kandelia candel, Rhizophora stylosa and Rhizophoraceae was 26, 20 and 16, respectively. After sequencing, 34 strains of endophytic fungi were found to have the same sequences. After removal of the repeated ones, 28 different strains of endophytic fungi were successfully isolated in the present study (Fig. 1). To further identify the 28 strains of endophytic fungi, the 18S rDNA was amplified by PCR. The results from agarose gel electrophoresis of the PCR products are shown in Fig. 2. As shown in Fig. 2, the size of the PCR products was from 500 to 750 bp, indicating that the 28 strains belonged to fungi. Compared with the sequences in the GenBank database, 28 different strains of endophytic fungi were successfully identified (Table I). Next, the phylogenetic trees were constructed using Mega 5.0 software. The phylogenetic trees of three types of mangroves, Kandelia candel, Rhizophora stylosa, and Rhizophoraceae, are shown in Figs. 35, respectively.

Table I.

Results of the identification of isolated endophytic fungi from mangrove plants.

Table I.

Results of the identification of isolated endophytic fungi from mangrove plants.

No.Strain codeSimilar strainsSources of similar strainsRates of similarity (%)Results of identificationNo.Strain codeSimilar strainsSources of similar strainsRates of similarity (%)Results of identification
  1zj-2Neofusicoccum australMangrove100Neofusicoccum sp.15zj-55Phomopsis sp. 125AS/TAnnona squamosa stem99Phomopsis sp.
zj-85(FJ037758.1) zj-10(GU066687.1)
zj-101 zj-56
zj-88
zj-70
  2zj-16Neofusicoccum australMyrtus communis99Neofusicoccum sp.16zj-11Phomopsis sp.139SD/SSpondias dulcis seed99Phomopsis sp.
zj-20E54ML (KF702388.1) zj-43(GU066698.1)
zj-35
zj-14
zj-67
  3zj-17Neofusicoccum sp. ZH4-E1Mangrove99Neofusicoccum sp.17gq-L12Phomopsis sp. ZZF08Excoecaria agallocha99Phomopsis sp.
zj-19(FJ037734.1) zj-23(EU236706.10)
zj-77 zj-81
  4zj-31Neofusicoccum sp.Unknown99Neofusicoccum sp.18gq-L17Phomopsis sp.Cocos nucifera95Phomopsis sp
ALG69 (KJ657714.1) zj-36Ac001 (JN857950.1)flower
  5zj-89Neofusicoccum parvumUnknown99Neofusicoccum sp.19zj-58Phomopsis sp. 20SO/L Saccharum99Phomopsis sp.
zj-93UY754 (EU080926.1) (GQ407098.1)officinarum leaf
  6zj-25Penicillium griseofulvuMangrove99Penicillium sp.20zj-9 LeptosphaerulinaUnknown96 Leptosphaerulina
zj-74091402 (EU664471.1) zj-37 chartarum
zj-86 zj-82(GQ254687.1)
zj-29
  7gq-L10Penicillium oxalicumUnknown99Penicillium sp.21zj-22Fungal endophyte sp.Unknown99Fungal endophyte sp.
NFML_CH42_88 zj-32 AiS7(EU054418.1)
(KM458819.1)
  8zj-80Penicillium citrinum MA-14Soil99Penicillium22zj-38Fungal sp. NIS3Tectona grandis bark91Fungal sp.
(HQ671192.1) citrinum zj-61(KF910769.1)
  9zj-4Pestalotiopsis sp. 1 AE-2013 Bradypus99Pestalotiopsis sp.23gq-L8 TrichodermaGinger99 Trichoderma
zj-45F4875 (KF746126.1) variegatus CHR2FC55Rhizosphere soil
(KJ591703.1)
10zj-48Pestalotiopsis FL21Huperzia serrata100Pestalotiopsis sp.24zj-30Hypocrea lixii SZMCUnknown99Hypocrea lixii
zj-100(KP689177.1) zj-9820858 (JX173851.1)
11gq-L1Pestalotiopsis sp. P18-11Unknown99Pestalotiopsis sp.25zj-33Arthrinium sp. 7Bamboo99Arthrinium sp.
zj-15(HQ262507.1) (HQ647335.1)
12gq-L5Pestalotiopsis sp. S149/2013Mushroom99Pestalotiopsis sp.26gq-L19Bipolaris papendorfiiPhaseolus vulgaris99 Bipolaris
gq-L3(KM041695.1) CMON22 (JQ753972.1)
13zj-50Pestalotiopsis DBT179/2013c2Mushroom99Pestalotiopsis27zj-42Fusarium oxysporumPigeon pea99Fusarium oxysporum
zj-83(KM041703.1) zj-68K9 (JF807396.1)
14zj-39Phomopsis sp. 89CN/FCocos nucifera99Phomopsis sp.28zj-79 STRI:ICBG-Panama: Saccharum99Unknown
zj-87(GU066658.1)flower TK1280 (KF436022.1)
zj-12
zj-64
Results of the MTT assay

To assess the antitumor activities of the 28 isolated strains of endophytic fungi, MTT assay was performed to observe the effects of these fungi on the growth of human lung cancer cells, A549 and NCI-H460. The results from the MTT assay are shown in Table II. As shown in Table II, 10 strains of endophytic fungi, including 4 Neofusicoccum sp. strains (zj-2, zj14, zj-17 and zj-67), 4 Phomopsis sp. strains (zj-12, zj-23, zj-36 and zj-70), 1 Leptosphaerulina sp. strain (zj-9) and 1 Penicillium sp. strain (zj-25), significantly inhibited the growth of lung cancer A549 and NCI-H460 cells. The average inhibitory rates of 10 strains in the A549 cells were 64.4, 59.5, 81.9, 43.9, 58.3, 56.2, 48.3, 42.4, 93.0 and 49.7%, respectively. The average inhibitory rates of 10 strains in the NCI-H460 cells were 41.2, 49.3, 82.7, 40.7, 53.9, 52.6, 56.8, 64.3%, 91.0 and 45.6%, respectively. The zj-9 and zj-17 strains showed a stronger inhibitory effects on both the A549 and NCI-H460 lung cancer cell lines. Particularly, zj-9 exhibited the strongest growth inhibitory activity in the two lung cancer cell lines.

Table II.

Results from the MTT assay.

Table II.

Results from the MTT assay.

No.Strain codeAverage Average inhibitory rates in A549 cells (%)Average Average inhibitory rates in NCI-H460 cells (%)No.Strain codeAverage Average inhibitory rates in A549 cells (%)Average Average inhibitory rates in NCI-H460 cells (%)
  1zj-264.441.215zj-559.40.14
zj-85 zj-10
zj-101 zj-56
zj-88
zj-70
  2zj-1656.914.416zj-1118.910.2
zj-20 zj-43
zj-35
zj-14
zj-67
  3zj-1781.982.717gq-L1216.037.2
zj-19 zj-23
zj-77 zj-81
  4zj-3143.613.718gq-L1748.356.8
zj-36
  5zj-8920.239.919zj-5816.039.4
zj-93
  6zj-2549.745.620zj-993.091.0
zj-74 zj-37
zj-86 zj-82
zj-29
  7gq-L1042.464.321zj-2247.732.1
zj-32
  8zj-807.611.522zj-3837.436.0
zj-61
  9zj-439.754.823gq-L815.117.9
zj-45
10zj-486.84.924zj-3064.97.5
zj-100 zj-98
11gq-L14.611.725zj-331.74.1
zj-15
12gq-L515.437.726gq-L1917.09.7
gq-L3
13zj-5017.424.227zj-4213.417.3
zj-83 zj-68
14zj-3921.447.728zj-7911.416.7
zj-87
zj-12
zj-64
Results of the in vitro angiogenesis assay

To further explore the underlying mechanism of the antitumor activity of the endophytic fungi, an in vitro angiogenesis assay was performed to observe the effects of endophytic fungi on the inhibition of lung cancer angiogenesis. In our previous studies, we found that human papillomavirus (HPV) type 16 E7 (HPV-16 E7) oncoprotein significantly enhanced lung cancer cell angiogenesis in vitro (26,27). Therefore, in the present study, we established an in vitro angiogenesis model induced by HPV-16 E7 oncoprotein. As shown in Fig. 6, HPV-16 E7 oncoprotein markedly stimulated microtubule formation (Fig. 6, image 2) as compared with the empty vector control (Fig. 6, image 1), which was in accordance with our previous studies (26,27), indicating that the model was successfully established. In the present study, we further found that endophytic fungi zj-14 (image 6), zj-17 (image 7) and zj-36 (image 10) markedly inhibited HPV-16 E7-stimulated microtubule formation (Fig. 6; P<0.01) in both A549 (Fig. 6A) and NCI-H460 (Fig. 6C) cells, which was further confirmed by total tube length (Fig. 6B and D; P<0.01). Additionally, endophytic fungus zj-23 (lane 8) inhibited HPV-16 E7-stimulated microtubule formation in the A549 cells (Fig. 6A and B; P<0.01) and endophytic fungus zj-12 (lane 5) inhibited HPV-16 E7-stimulated microtubule formation in the NCI-H460 cells (Fig. 6C and D; P<0.05).

Discussion

In the present study, we isolated 26, 20 and 16 strains of endophytic fungi from three types of mangrove plants, Kandelia candel, Rhizophora stylosa and Rhizophoraceae, respectively. There are a variety of endophytic fungi that may be isolated from one type of mangrove plant, but one to three endophytic fungi are dominant. We found that the dominant endophytic fungi of Kandelia candel, Rhizophora stylosa and Rhizophoraceae were Pestalotiopsis sp. (42.3%), Pestalotiopsis sp. (20%) and Phomopsis sp. (43.8%), respectively. Notably, Neofusicoccum sp. (31.3%) was also found to be a major advantage endophytic fungi isolated from Rhizophoraceae in addition to Phomopsis sp.

A growing body of evidence indicates that multiple bioactive compounds isolated from mangrove-derived endophytic fungi inhibit the growth of cancer cells (1518). A new sesquiterpene named botryosphaerin F from the mangrove fungus Aspergillus terreus (no. GX7-3B) was reported to inhibit the growth of human breast cancer MCF-7 and leukemia HL-60 cells with IC50 values of 4.49 and 3.43 µM, respectively (15). Five highly oxygenated chromones, rhytidchromones A-E, were isolated from the culture broth of a mangrove-derived endophytic fungus, Rhytidhysteron rufulum, and all compounds, except for rhytidchromone D, displayed cytotoxicity against Kato-3 cancer cells with IC50 values ranging from 16.0 to 23.3 µM, while rhytidchromones A and C showed activity against breast cancer MCF-7 cells with IC50 values of 19.3 and 17.7 µM, respectively (16). Four new lasiodiplodins (14) were isolated from a mangrove endophytic fungus, Lasiodiplodia sp. 318#., and compound 4 exhibited moderate cytotoxic activities against human cancer lines THP1, MDA-MB-435, A549, HepG2 and HCT-116 (17). Two of the three new resveratrol derivatives, namely resveratrodehydes, isolated from the mangrove endophytic fungus Alternaria sp. R6, were found to exhibit cytotoxic activities against human breast cancer cell line MDA-MB-435 and human colon cancer cell line HCT-116s (IC50 <10 µM) (18). Additionally, Tao et al (28) separated 87 compounds from mangrove endophytic fungus in Southern China, of which 14% of the compounds had antitumor activity. In the present study, we found that 10 strains, including Neofusicoccum sp. 4 strains (zj-2, zj14, zj-17 and zj-67), Phomopsis sp. 4 strains (zj-12, zj-23, zj-36 and zj-70), Leptosphaerulina sp. 1 strain (zj-9) and Penicillium sp. 1 strain (zj-25), significantly inhibited the growth of the lung cancer cells, A549 and NCI-H460.

Angiogenesis plays a key role in cancer invasion and metastasis. Thus, inhibition of angiogenesis is an effective strategy for cancer prevention (25,29,30). At present, multiple natural compounds have been reported in in vitro and in vivo experiments to inhibit angiogenesis, and most of them from terrestrial plants (2933). Recently, the screening of marine medicines from the sea and its surroundings has also attracted increased attention. A number of angiogenic inhibitors from marine organisms have been found (34,35). Moreover, anti-angiogenic drugs can also be isolated from endophytic fungi (3638). Altersolanol, isolated from an Alternaria sp. endophytic fungus, was reported to show promising anti-angiogenic activity ex vivo, in vitro and in vivo by the suppression of proliferation, tube formation and migration (36). The phenolic compounds, isolated from an endophytic fungus Coccomyces proteae collected from a Costa Rican rainforest, were reported to have anti-angiogenic activity via inhibition of capillary morphogenesis gene protein 2 (CMG2) (37). Particularly, toluquinol, isolated from marine fungus secondary metabolites, was also demonstrated to inhibit angiogenesis both in vitro and in vivo partly by the suppression of VEGF and FGF-induced Akt activation (38). In the present study, we established an HPV-16 E7 oncoprotein-induced lung cancer angiogenic model according to our previous findings (26,27), and further observed the effects of endophytic fungi which have high inhibitory effect on angiogenesis in vitro. We found that zj-14, zj-17 and zj-36 endophytic fungi significantly inhibited lung cancer angiogenesis in vitro. Notably, strain zj-9 was found to have the strongest inhibitory effect on the growth of lung cancer cells, but it did not exhibit anti-angiogenic activity, indicating that strain zj-9 may have cell cytotoxicity but not anti-angiogenic activity, and this issue warrants further study.

Acknowledgements

The present study was supported by grants from the National Natural Science Foundation of China (no. 81372511) (to X.T.), the Guangdong Provincial Department of Science and Technology (Research and Development of Industrial Technology in Guangdong Province) (no. 2013B031100002) (to X.T.), and the Zhanjiang Municipal Governmental Specific Financial Fund Allocated for Competitive Scientific and Technological Projects (no. 2012C0303-56) (to X.T.).

References

1 

Campos FF, Junior PA Sales, Romanha AJ, Araújo MS, Siqueira EP, Resende JM, Alves TM, Martins-Filho OA, Santos VL, Rosa CA, et al: Bioactive endophytic fungi isolated from Caesalpinia echinata Lam. (Brazilwood) and identification of beauvericin as a trypanocidal metabolite from Fusarium sp. Mem Inst Oswaldo Cruz. 110:65–74. 2015. View Article : Google Scholar : PubMed/NCBI

2 

Sun ZH, Liang FL, Chen YC, Liu HX, Li HH and Zhang WM: Two new xyloketals from the endophytic fungus Endomelanconiopsis endophytica derived from medicinal plant Ficus hirta. J Asian Nat Prod Res. 18:1036–1041. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Sun W, Chen X, Tong Q, Zhu H, He Y, Lei L, Xue Y, Yao G, Luo Z, Wang J, et al: Novel small molecule 11β-HSD1 inhibitor from the endophytic fungus Penicillium commune. Sci Rep. 6:264182016. View Article : Google Scholar : PubMed/NCBI

4 

Xu DB, Ye WW, Han Y, Deng ZX and Hong K: Natural products from mangrove actinomycetes. Mar Drugs. 12:2590–2613. 2014. View Article : Google Scholar : PubMed/NCBI

5 

Debbab A, Aly AH, Lin WH and Proksch P: Bioactive compounds from marine bacteria and fungi. Microb Biotechnol. 3:544–563. 2010. View Article : Google Scholar : PubMed/NCBI

6 

Wang J, Ding W, Wang R, Du Y, Liu H, Kong X and Li C: Identification and bioactivity of compounds from the mangrove endophytic fungus Alternaria sp. Mar Drugs. 13:4492–4504. 2015. View Article : Google Scholar : PubMed/NCBI

7 

Cui H, Liu Y, Nie Y, Liu Z, Chen S, Zhang Z, Lu Y, He L, Huang X and She Z: Polyketides from the mangrove-derived endophytic fungus Nectria sp. HN001 and their α-glucosidase inhibitory activity. Mar Drugs. 14:pii: E862016. View Article : Google Scholar

8 

Ding B, Wang Z, Huang X, Liu Y, Chen W and She Z: Bioactive α-pyrone meroterpenoids from mangrove endophytic fungus Penicillium sp. Nat Prod Res. Apr 11;1–8. 2016.(Epub ahead of print).

9 

Liu Y, Chen S, Liu Z, Lu Y, Xia G, Liu H, He L and She Z: Bioactive metabolites from mangrove endophytic fungus Aspergillus sp. 16-5B. Mar Drugs. 13:3091–3102. 2015. View Article : Google Scholar : PubMed/NCBI

10 

Bai ZQ, Lin X, Wang Y, Wang J, Zhou X, Yang B, Liu J, Yang X, Wang Y and Liu Y: New phenyl derivatives from endophytic fungus Aspergillus flavipes AIL8 derived of mangrove plant Acanthus ilicifolius. Fitoterapia. 95:194–202. 2014. View Article : Google Scholar : PubMed/NCBI

11 

Zhou XM, Zheng CJ, Chen GY, Song XP, Han CR, Tang XZ, Liu RJ and Ren LL: Two new stemphol sulfates from the mangrove endophytic fungus Stemphylium sp. 33231. J Antibiot. 68:501–503. 2015. View Article : Google Scholar : PubMed/NCBI

12 

Meng LH, Zhang P, Li XM and Wang BG: Penicibrocazines A-E, five new sulfide diketopiperazines from the marine-derived endophytic fungus Penicillium brocae. Mar Drugs. 13:276–287. 2015. View Article : Google Scholar : PubMed/NCBI

13 

Lv F, Daletos G, Lin W and Proksch P: Two new cyclic depsipeptides from the endophytic fungus Fusarium sp. Nat Prod Commun. 10:1667–1670. 2015.PubMed/NCBI

14 

Wang M, Zhang W, Xu W, Shen Y and Du L: Optimization of genome shuffling for high-yield production of the antitumor deacetylmycoepoxydiene in an endophytic fungus of mangrove plants. Appl Microbiol Biotechnol. 100:7491–7498. 2016. View Article : Google Scholar : PubMed/NCBI

15 

Deng C, Huang C, Wu Q, Pang J and Lin Y: A new sesquiterpene from the mangrove endophytic fungus Aspergillus terreus (No. GX7-3B). Nat Prod Res. 27:1882–1887. 2013. View Article : Google Scholar : PubMed/NCBI

16 

Chokpaiboon S, Choodej S, Boonyuen N, Teerawatananond T and Pudhom K: Highly oxygenated chromones from mangrove-derived endophytic fungus Rhytidhysteron rufulum. Phytochemistry. 122:172–177. 2016. View Article : Google Scholar : PubMed/NCBI

17 

Li J, Xue Y, Yuan J, Lu Y, Zhu X, Lin Y and Liu L: Lasiodiplodins from mangrove endophytic fungus Lasiodiplodia sp. 318. Nat Prod Res. 30:755–760. 2016. View Article : Google Scholar : PubMed/NCBI

18 

Wang J, Cox DG, Ding W, Huang G, Lin Y and Li C: Three new resveratrol derivatives from the mangrove endophytic fungus Alternaria sp. Mar Drugs. 12:2840–2850. 2014. View Article : Google Scholar : PubMed/NCBI

19 

Huang L, Zhang T, Li S, Duan J, Ye F, Li H, She Z, Gao G and Yang X: Anthraquinone G503 induces apoptosis in gastric cancer cells through the mitochondrial pathway. PLoS One. 9:e1082862014. View Article : Google Scholar : PubMed/NCBI

20 

Chen WL, Turlova E, Sun CL, Kim JS, Huang S, Zhong X, Guan YY, Wang GL, Rutka JT, Feng ZP, et al: Xyloketal B suppresses glioblastoma cell proliferation and migration in vitro through inhibiting TRPM7-regulated PI3K/Akt and MEK/ERK signaling pathways. Mar Drugs. 13:2505–2525. 2015. View Article : Google Scholar : PubMed/NCBI

21 

Wang X, Tan T, Mao ZG, Lei N, Wang ZM, Hu B, Chen ZY, She ZG, Zhu YH and Wang HJ: The marine metabolite SZ-685C induces apoptosis in primary human nonfunctioning pituitary adenoma cells by inhibition of the Akt pathway in vitro. Mar Drugs. 13:1569–1580. 2015. View Article : Google Scholar : PubMed/NCBI

22 

Xie G, Zhu X, Li Q, Gu M, He Z, Wu J, Li J, Lin Y, Li M, She Z, et al: SZ-685C, a marine anthraquinone, is a potent inducer of apoptosis with anticancer activity by suppression of the Akt/FOXO pathway. Br J Pharmacol. 159:689–697. 2010. View Article : Google Scholar : PubMed/NCBI

23 

Wang D, Wang S, Liu Q, Wang M, Wang C and Yang H: SZ-685C exhibits potent anticancer activity in both radiosensitive and radioresistant NPC cells through the miR-205-PTEN-Akt pathway. Oncol Rep. 29:2341–2347. 2013.PubMed/NCBI

24 

Zhu X, He Z, Wu J, Yuan J, Wen W, Hu Y, Jiang Y, Lin C, Zhang Q, Lin M, et al: A marine anthraquinone SZ-685C overrides adriamycin-resistance in breast cancer cells through suppressing Akt signaling. Mar Drugs. 10:694–711. 2012. View Article : Google Scholar : PubMed/NCBI

25 

Folkman J: Role of angiogenesis in tumor growth and metastasis. Semin Oncol. 29 Suppl 16:S15–S18. 2002. View Article : Google Scholar

26 

Li G, He L, Zhang E, Shi J, Zhang Q, Le AD, Zhou K and Tang X: Overexpression of human papillomavirus (HPV) type 16 oncoproteins promotes angiogenesis via enhancing HIF-1α and VEGF expression in non-small cell lung cancer cells. Cancer Lett. 311:160–170. 2011. View Article : Google Scholar : PubMed/NCBI

27 

Zhang E, Feng X, Liu F, Zhang P, Liang J and Tang X: Roles of PI3K/Akt and c-Jun signaling pathways in human papillomavirus type 16 oncoprotein-induced HIF-1α, VEGF, and IL-8 expression and in vitro angiogenesis in non-small cell lung cancer cells. PLoS One. 9:e1034402014. View Article : Google Scholar : PubMed/NCBI

28 

Tao YW, Lin YC, She Z-G, Lin MT, Chen PX and Zhang JY: Anticancer activity and mechanism investigation of beauvericin isolated from secondary metabolites of the mangrove endophytic fungi. Anticancer Agents Med Chem. 15:258–266. 2015. View Article : Google Scholar : PubMed/NCBI

29 

Shi J, Liu F, Zhang W, Liu X, Lin B and Tang X: Epigallocatechin-3-gallate inhibits nicotine-induced migration and invasion by the suppression of angiogenesis and epithelial-mesenchymal transition in non-small cell lung cancer cells. Oncol Rep. 33:2972–2980. 2015.PubMed/NCBI

30 

He L, Zhang E, Shi J, Li X, Zhou K, Zhang Q, Le AD and Tang X: (−)-Epigallocatechin-3-gallate inhibits human papillomavirus (HPV)-16 oncoprotein-induced angiogenesis in non-small cell lung cancer cells by targeting HIF-1α. Cancer Chemother Pharmacol. 71:713–725. 2013. View Article : Google Scholar : PubMed/NCBI

31 

Lee J, Yi JM, Kim H, Lee YJ, Park JS, Bang OS and Kim NS: Cytochalasin H, an active anti-angiogenic constituent of the ethanol extract of Gleditsia sinensis thorns. Biol Pharm Bull. 37:6–12. 2014. View Article : Google Scholar : PubMed/NCBI

32 

Lee SR, Park JY, Yu JS, Lee SO, Ryu JY, Choi SZ, Kang KS, Yamabe N and Kim KH: Odisolane, a novel oxolane derivative, and antiangiogenic constituents from the fruits of mulberry (Morus alba L.). J Agric Food Chem. 64:3804–3809. 2016. View Article : Google Scholar : PubMed/NCBI

33 

Park JY, Shin MS, Kim SN, Kim HY, Kim KH, Shin KS and Kang KS: Polysaccharides from Korean Citrus hallabong peels inhibit angiogenesis and breast cancer cell migration. Int J Biol Macromol. 85:522–529. 2016. View Article : Google Scholar : PubMed/NCBI

34 

Goey AK, Chau CH, Sissung TM, Cook KM, Venzon DJ, Castro A, Ransom TR, Henrich CJ, McKee TC, McMahon JB, et al: Screening and biological effects of marine pyrroloiminoquinone alkaloids: Potential inhibitors of the HIF-1α/p300 interaction. J Nat Prod. 79:1267–1275. 2016. View Article : Google Scholar : PubMed/NCBI

35 

Baharara J, Amini E and Mousavi M: The anti-proliferative and anti-angiogenic effect of the methanol extract from brittle star. Rep Biochem Mol Biol. 3:68–75. 2015.PubMed/NCBI

36 

Pompeng P, Sommit D, Sriubolmas N, Ngamrojanavanich N, Matsubara K and Pudhom K: Antiangiogenetic effects of anthranoids from Alternaria sp., an endophytic fungus in a Thai medicinal plant Erythrina variegata. Phytomedicine. 20:918–922. 2013. View Article : Google Scholar : PubMed/NCBI

37 

Cao S, Cryan L, Habeshian KA, Murillo C, Tamayo-Castillo G, Rogers MS and Clardy J: Phenolic compounds as antiangiogenic CMG2 inhibitors from Costa Rican endophytic fungi. Bioorg Med Chem Lett. 22:5885–5888. 2012. View Article : Google Scholar : PubMed/NCBI

38 

García-Caballero M, Marí-Beffa M, Cañedo L, Medina MÁ and Quesada AR: Toluquinol, a marine fungus metabolite, is a new angiosuppresor that interferes with the Akt pathway. Biochem Pharmacol. 85:1727–1740. 2013. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

March-2017
Volume 37 Issue 3

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Liu X, Wu X, Ma Y, Zhang W, Hu L, Feng X, Li X and Tang X: Endophytic fungi from mangrove inhibit lung cancer cell growth and angiogenesis in vitro Corrigendum in /10.3892/or.2023.8608. Oncol Rep 37: 1793-1803, 2017
APA
Liu, X., Wu, X., Ma, Y., Zhang, W., Hu, L., Feng, X. ... Tang, X. (2017). Endophytic fungi from mangrove inhibit lung cancer cell growth and angiogenesis in vitro Corrigendum in /10.3892/or.2023.8608. Oncology Reports, 37, 1793-1803. https://doi.org/10.3892/or.2017.5366
MLA
Liu, X., Wu, X., Ma, Y., Zhang, W., Hu, L., Feng, X., Li, X., Tang, X."Endophytic fungi from mangrove inhibit lung cancer cell growth and angiogenesis in vitro Corrigendum in /10.3892/or.2023.8608". Oncology Reports 37.3 (2017): 1793-1803.
Chicago
Liu, X., Wu, X., Ma, Y., Zhang, W., Hu, L., Feng, X., Li, X., Tang, X."Endophytic fungi from mangrove inhibit lung cancer cell growth and angiogenesis in vitro Corrigendum in /10.3892/or.2023.8608". Oncology Reports 37, no. 3 (2017): 1793-1803. https://doi.org/10.3892/or.2017.5366