Effect of liposome‑mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells

  • Authors:
    • Haijing Deng
    • Xuemin Gao
    • Haibing Peng
    • Jin Wang
    • Xiaoli Hou
    • Hong Xu
    • Fang Yang
  • View Affiliations

  • Published online on: March 14, 2018     https://doi.org/10.3892/mmr.2018.8744
  • Pages: 7319-7324
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

To investigate the effect of liposome Lipofectamine® 2000‑mediated HSP27 plasmid transfection in A549 human alveolar type II epithelial cell line on collagen synthesis during transforming growth factor‑β1 (TGF‑β1)‑induced type II epithelial cell transition to myofibroblasts. Cells were transfected with varying ratios of the Lipofectamine® 2000‑mediated heat shock protein 27 (HSP27) plasmid and the transfection efficiency was determined using flow cytometry. The maximum transfection efficacy was confirmed by laser confocal microscopy. HSP gene expression and the most efficient HSP27 plasmid were determined using reverse transcription‑quantitative polymerase chain reaction. Western blot analysis was used to examine HSP27 and collagen expression levels. With a transfection efficiency of 83%, the 8 µg:20 µl ratio of liposome: Plasmid had the highest transfection levels. Among the four different interference sequences in the HSP27 plasmid, the D sequence had the highest interference effect with 70% silencing of the HSP27 gene. The expression of type I and III collagen in TGF‑β1‑induced transition of A549 human alveolar type II epithelial cell line to myofibroblasts was significantly downregulated by the successful transfection with HSP27‑interfering plasmid. The expression of type I and III collagen in the TGF‑β1‑induced transition of A549 cells to myofibroblasts was significantly downregulated by transfection of A549 cells with HSP27 plasmid D‑interfering sequence and optimal ratio of Lipofectamine® 2000 and HSP27 plasmid.

Introduction

Heat shock protein 27 (HSP27) is a differential protein isolated from silicosis tissue (1). HSP27 belongs to a small molecular weight HSP family, whose proteins contain a conservative C-terminal domain, termed the α-lens protein domain. Previous studies revealed that HSP27 may antagonize oxidative stress, inhibit cell apoptosis and act as a chaperone in inflammatory reactions, cell signal transduction, cell differentiation, and proliferation by regulating actin cytoskeleton structure (27). HSP27 is involved in the epithelial-mesenchymal transition (EMT) in cancer (8) and in the development and progression of organ fibrosis (9). However, previous studies have also revealed that HSP27 may exert dual regulation during EMT in organ fibrosis (9,10). Transforming growth factor-β1 (TGF-β1) is an important transforming growth factor associated with fibrosis in vitro and in vivo (11,12). A previous study by the authors revealed that TGF-β1 may induce A549 human alveolar type II epithelial cells to differentiate into myofibroblasts (13). In order to investigate the effect of HSP27 on the differentiation of A549 human alveolar type II epithelial cell line into myofibroblasts and collagen synthesis, the current study used liposome transfection to transfect A549 human alveolar type II epithelial cell line and determined the optimal liposome:plasmid ratio and the best interference sequence. Lipofectamine® 2000 was used in the current study as the protocol was established, safe and reliable with high transfection efficacy. Following successful transfection of the HSP27 interfering plasmid, the effect on collagen expression during TGF-β1-induced differentiation of A549 human alveolar type II epithelial cells into myofibroblast was examined.

Materials and methods

Reagents and materials

The following reagents were used in the study: A549 human alveolar type II epithelial cell line derived from human lung carcinoma (cat. no. TCHu150, Cell Bank of Chinese Academy of Science, Shanghai, China); high-glucose Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA); 10% fetal bovine serum (A15-101, PAA, New Bedford, Massachusetts, USA; www.openfos.com/supply/3298452-PAA-LAB-in-New-Bedford-MA/); TGF-β1 (cat. no. 100-21; Peprotech, Inc., Rocky Hill, NJ, USA), mouse anti-HSP27 monoclonal antibody (cat. no. ab114067; Abcam, Cambridge, UK), rabbit polyclonal anti-type I/type III collagen antibody (cat. nos. BA0325/0326, GeneTex, Inc., Irvine, CA, USA), Platinum® SYBR®-Green qPCR SuperMix-UDG kit (cat. no. C11733038, Invitrogen; Thermo Fisher Scientific, Inc.), Lipofectamine® 2000 transfection reagent (1.5 ml/U; Invitrogen; Thermo Fisher Scientific, Inc.); HSP27-interfering kit including fragments A, B, C and D (Shanghai GenePharma Co., Ltd., Shanghai, China); Opti-MEM medium (500 ml/bottle, Gibco; Thermo Fisher Scientific, Inc.); and polyclonal anti-GAPDH antibody (cat. no. sc-25778; Santa Cruz Biotechnology, Inc., Dallas, TX, USA).

Cell culture and experimental groups

Specific HSP27 gene fragment was introduced into A549 cells by transfection. Cells were divided into three groups: i) Blank plasmid control group; ii) blank plasmid + TGF-β1 group; and iii) HSP27-interfering plasmid + TGF-β1 group.

Transfection efficacy determined by flow cytometry

Cells were seeded into 10 cm diameter dishes and transfected at 90–100% confluency. Initially, two aliquots of 3 ml Opti-MEM medium was added into two 5 ml Eppendorf micro test tubes, Lipofectamine® 2000 and HSP27 plasmid were added individually into the two tubes and mixed with OPTI-MEM medium. A series of Lipofectamine® 2000 volumes (20, 25, 30, 20, and 20 µl) and HSP27 plasmid (8, 8, 8, 9 and 10 µg) were correspondingly tested. The reagents were individually incubated at 37°C for 10 min prior being mixed and incubated at 37°C for an additional 20 min. The transfection mixture was added into dishes with 15 ml OPTI-MEM and mixed by gentle shaking. After 72 h incubation at 37°C, cells were trypsinized with 0.25% trypsin, harvested, and centrifuged at 201 × g on 4°C for 5 min. Transfection efficacy of each combination of Lipofectamine® 2000 and plasmid was determined using fluorescent intensity via flow cytometry.

Examination by laser confocal microscopy

Cells were seeded at 6×103 cells/well in the small dish for confocal observation. HSP27 plasmid and Lipofectamine® 2000 were added at 8 µg:20 µl ratio for transfection as aforementioned. The fluorescence marker green fluorescent protein (GFP) in the plasmid cells were observed under laser confocal microscope at a magnification of ×800.

Screen optimal interfering HSP27 plasmid by reverse transcription-quantitative polymerase chain reaction (RT-qPCR)

Cells were seeded at a density of 6×103 cells/well in 6-well plate and transfected when cells reached 90–100% confluency. For transfection, 500 µl OPTI-MEM medium was added into 1.5 ml Eppendorf tube and 12 tubes were prepared for 6 transfection groups, including 4 different HSP27 interfering sequences, HSP27 positive control and negative control plasmids; 5 µl Lipofectamine® 2000 transfection reagent or the plasmid was mixed with OPTI-MEM and incubated for 10 min. Then the transfection reagent mixture was added into plasmid/OPTI-MEM mixture and incubated at 37°C for 20 min. RNA was isolated with TRIzol (Invitrogen, Thermo Fisher Scientific, Inc.) after 72 h and the concentration and purity of total RNA was measured with deionized water as control by micro nucleic acid analyzer. The primer of target gene was designed and synthesized (Invitrogen; Thermo Fisher Scientific, Inc.). HSP27: Forward, 5′-GCTTCACGCGGAAATACACG-3′ and reverse, 5′-GTGATCTCGTTGGACTGCGT-3′; β-actin forward, 5′-GTCACCTTCACCGTTCCAGTTTT-3′ and reverse, 5′-CTTAGTTGCGTTACACCCTTTCTT-3′. qPCR amplification was performed as follows: Pre-denaturation at 95°C for 15 sec followed by 40 cycles at 60°C for 20 sec. And data were analyzed to determine the optimal HSP27 interfering sequence by the method of ∆∆Cq (14).

Immunoblotting

Protein was isolated from cells with RIPA (Beijing Solarbio Science & Technology Co., Ltd. Beijing, China) following the designated transfection period. After determining the protein concentration by BCA assay, 50 µl samples per lane were loaded on 10% SDS-PAGE. The blots were subjected to standard transfer methods and PVDF membranes were blocked with 10% BSA (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany), and then incubated with primary antibodies (HSP27, type I and III collagen all at 1:200 dilution and GAPDH at 1:100 dilution) at 4°C overnight. The membranes were incubated with goat anti-rabbit IgG-AP (BA1011; Boster Biological Technology, Pleasanton, CA, USA) and goat anti-mouse IgG-AP (BA1010; Boster Biological Technology) secondary antibodies diluted at 1:5,000 at 37°C for 2 h and then with RCIP/NBT chromogenic reagent for 1 min. The optical density of protein band was determined by ImageJ software (National Institutes of Health, Bethesda, MD, USA). Relative expression of protein was calculated by the ratio of optical density of target protein band to internal control band. Statistical analysis was performed on relative protein expression.

Statistical analysis

SPSS version 17.0 (SPSS, Inc., Chicago, IL, USA) was used for statistical analysis which included Bartlett's, Brown-Forsythe and Levene's tests, to test for equal variance followed by Student-Newman-Keuls multiple-range test and one-way analysis of variance was used to compare differences between groups.

Results

Transfection efficacy of Lipofectamine® 2000 and HSP27 assessed by flow cytometry

Five different combinations of HSP27 plasmid and transfection reagent Lipofectamine® 2000 were assessed, including 8 µg:20 µl; 8 µg:25 µl; 8 µg:30 µl; 9 µg:20 µl; 10 µg:20 µl. Flow cytometry data indicated in Fig. 1 revealed that transfection efficacy was 83, 76, 76, 70, 74%, respectively. The 8 µg:20 µl ratio led to a 83% transfection efficiency and was considered to be the optimal liposome to plasmid ratio and used for subsequent experiments.

Laser scanning confocal microscope detection for the optimal ratio of Lipofectamine® 2000 and HSP27 with best transfection efficacy

Flow cytometry results were confirmed with microscopy and after successful transfection at 8 µg:20 µl of HSP27: Lipofectamine® 2000, GFP positive expression was visible in the cytoplasm (Fig. 2).

Optimal interfering HSP27 plasmid identified using RT-qPCR

Data from the RT-qPCR are presented in Fig. 3 and indicated that the interference effect of the four different sequences was 48, 55, 59 and 30% with respect to the positive control group and 48, 54, 58 and 30% relative to negative control group. The fourth interfering sequence (D) achieved the most efficient gene silencing of HSP27 with 70% of the HSP27 gene being silenced, which was statistically significant compared with positive control and negative control group.

Transfection efficacy and the expression of type I and III collagen

Data from western blot experiments revealed that compared with the HSP27 negative control group, the expression of HSP27 and phosphorylated HSP27 in transfection of HSP27-interfering plasmid group was downregulated (Fig. 4; P<0.05). The difference was statistically significant compared with the transfection of GAPDH in the positive control group, the expression of GAPDH was significantly upregulated in transfected with HSP27 interfering plasmid and the HSP27 negative control group (P<0.05). This indicated that the transfection of HSP27 interference plasmid was successful. Additionally, findings from the western blot analysis revealed that compared with the blank plasmid transfection group, the expression of HSP27 and pHSP27 increased, which was accompanied by the increased expression of type I and III collagen in blank plasmid group, which was induced with TGF-β1 (Fig. 5), which was statistically significant (P<0.05). The expression of HSP27 and pHSP27 was significantly downregulated along with the reduced expression of type I and III collagen in the TGF-β1-induced HSP27-interfering plasmid group, compared with the TGF-β1-induced blank plasmid transfection group (P<0.05).

Discussion

Cationic liposome is a promising non-virus gene vehicle (15,16), whose membrane property confers various advantages in application terms of therapy and study of gene transfection, including maintaining the biological activity of genes, protecting genes from degradation by lysosomes. It is also non-immunogenic, easy to apply, with high repeatability and it is naturally degraded (1725). The current study used Lipofectamine® 2000 to transfect A549 human alveolar type II epithelial cell line with HSP27-interfering plasmid; the procedure was easy, safe and reliable with high transfection efficacy, and met the requirement of subsequent experiments. Therefore, this method provides a reference in the application of Lipofectamine® 2000-mediated transfection of other cell lines.

HSP27 is a small molecular weight protein in heat shock protein family (26). Previous studies have previously reported that HSP27 is involved in the tumor cell EMT (2732). In breast cancer, tumor cells that undergo EMT exhibit a high expression of HSP27 protein, which indicated that HSP27 protein may be closely associated with breast cancer cell EMT and may be involved in tumor migration and drug resistance (32). When MET-5A pleural mesothelial cells were stimulated in vitro with TGF-β1, the expression of HSP27 was significantly upregulated and accompanied by the increased expression of α-smooth muscle actin (α-SMA) and reduced the expression of E-cadherin, whereas cell morphology changed from flat cobblestones to long spindle-shapes. When HSP27-specific inhibitor OGX-427 (antisense oligonucleotides) or HSP27 siRNA transfection were used the increased expression of HSP27 and α-SMA protein by TGF-β stimulation was effectively reduced (9). Additionally, the downregulation of E-cadherin expression was also inhibited and cell morphology was maintained as flat or round-like. This indicated that HSP27 may be important for fibrosis and involved in pleural mesothelial cell EMT (9). In NRK52E tubular epithelial cells, TGF-β1 stimulation led to the upregulation of HSP27 expression, accompanied by the upregulation of α-SMA and vimentin, as well as fibronectin expression and led to downregulation of E-cadherin expression. This suggested that HSP27 has a protective role in tubular epithelial cell EMT (10). Additionally, HSP27 had a protective effect on myocardial cells in ischemic heart disease and adriamycin-induced myocardial injury due to this anti-apoptotic effect (33,34). Therefeore, it is important to investigate the effect of HSP27 on A549 human alveolar type II epithelial cell EMT. Extracellular matrix (ECM) deposition is a direct cause of fibrogenesis (3539). Type I and III collagen are essential ECM components that reflect the differentiation extent of myofibroblasts (4043). In our previous studies, we found that 5 ng/ml TGF-β1 was able to successfully induce the transition of A549 human alveolar type II epithelial cells to myofibroblasts, accompanied with upregulation of type I and III collagen expression (13). In A549 cells transfected with HSP27-interfering plasmid, the expression of type I and III collagen was downregulated by TGF-β1 stimulation, which suggested that HSP27 is a factor that may induce fibrosis in the transition of TGF-β1-induced A549 human alveolar type II epithelial cells to myofibroblasts. The present study provided the experimental basis and theoretical evidence for further investigation of the effect of HSP27 in vivo on EMT in silicosis or fibrosis.

Acknowledgements

Not applicable.

Funding

The present study was supported by funding from the National Natural Science Foundation of China (grant no. 81302395) and Science and Technology Research Projects in the Colleges and Universities of Hebei Province (grant no. QN2016147).

Availability of data and materials

The analyzed data generated during the study are available from the corresponding author on reasonable request.

Authors' contributions

FY conceived and designed the experiments. HJD, XMG, HBP and XLH performed the experiments. HJD, HX and JW analyzed the data. FY wrote the manuscript and HJD provided support on reagents/materials/analysis tools.

Ethics approval and consent to participate

A549 human alveolar type II epithelial cell line derived from human lung carcinoma purchased from Cell Bank of Chinese Academy of Science, Shanghai, China which was received ethics approval. The manuscript involves no other human participants, human data or human tissue.

Consent for publication

All authors consent for publication.

Competing interests

The authors declare that they have no competing interests.

References

1 

Xu H, Xue X, Du S, Li S, Sun Y, Yuan Y, Deng H, Wei Z, Wang R and Yang F: Comparative proteomic analysis on anti-fibrotic effect of N-acetyl-seryl-aspartyl-lysyl-proline in rats with silicosis. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 32:561–567. 2014.(In Chinese). PubMed/NCBI

2 

Tang S, Chen H, Cheng Y, Nasir MA, Kemper N and Bao E: Expression profiles of heat shock protein 27 and αB-crystallin and their effects on heat-stressed rat myocardial cells in vitro and in vivo. Mol Med Rep. 13:1633–1638. 2016. View Article : Google Scholar : PubMed/NCBI

3 

Song IS, Kang SS, Kim ES, Park HM, Choi CY, Tchah H and Kim JY: Heat shock protein 27 phosphorylation is involved in epithelial cell apoptosis as well as epithelial migration during corneal epithelial wound healing. Exp Eye Res. 118:36–41. 2014. View Article : Google Scholar : PubMed/NCBI

4 

Zhao M, Ding JX, Zeng K, Zhao J, Shen F, Yin YX and Chen Q: Heat shock protein 27: A potential biomarker of peritoneal metastasis in epithelial ovarian cancer? Tumour Biol. 35:1051–1056. 2014. View Article : Google Scholar : PubMed/NCBI

5 

Zembron-Lacny A, Ziemann E, Zurek P and Hübner-Wozniak E: Heat shock protein 27 response to wrestling training in relation to the muscle damage and inflammation. J Strength Cond Res. 31:1221–1228. 2017. View Article : Google Scholar : PubMed/NCBI

6 

Wang S, Yang S, Vlantis AC, Liu SY, Ng EK, Chan AB, Wu J, Du J, Wei W, Liu X, et al: Expression of antioxidant molecules and heat shock protein 27 in thyroid tumors. J Cell Biochem. 117:2473–2481. 2016. View Article : Google Scholar : PubMed/NCBI

7 

Carper SW, Rocheleau TA, Cimino D and Storm FK: Heat shock protein 27 stimulates recovery of RNA and protein synthesis following a heat shock. J Cell Biochem. 66:153–164. 1997. View Article : Google Scholar : PubMed/NCBI

8 

Schweiger T, Nikolowsky C, Starlinger P, Traxler D, Zimmermann M, Birner P, Hegedüs B, Dome B, Bergmann M, Mildner M, et al: Stromal expression of heat-shock protein 27 is associated with worse clinical outcome in patients with colorectal cancer lung metastases. PLoS One. 10:e01207242015. View Article : Google Scholar : PubMed/NCBI

9 

Wettstein G, Bellaye PS, Kolb M, Hammann A, Crestani B, Soler P, Marchal-Somme J, Hazoume A, Gauldie J, Gunther A, et al: Inhibition of HSP27 blocks fibrosis development and EMT features by promoting Snail degradation. FASEB J. 27:1549–1560. 2013. View Article : Google Scholar : PubMed/NCBI

10 

Vidyasagar A, Reese S, Acun Z, Hullett D and Djamali A: HSP27 is involved in the pathogenesis of kidney tubulointerstitial fibrosis. Am J Physiol Renal Physiol. 295:F707–F716. 2008. View Article : Google Scholar : PubMed/NCBI

11 

Sun YC, Liang Q, Qian KL, Xiao L, Liu Q and Shi XF: Effect of TGF-b1 siRNA-mediated silencing on Smad proteins in hepatic fibrosis rats. Zhonghua Gan Zang Bing Za Zhi. 20:289–293. 2012.(In Chinese). PubMed/NCBI

12 

Cheng J, Wang M, Ma H, Li H, Ren J and Wang R: Adiponectin inhibits oxidative stress and modulates TGF-b1 and COL-1 expression via the AMPK pathway in HSC-T6 cells. Zhonghua Gan Zang Bing Za Zhi. 23:69–72. 2015.(In Chinese). PubMed/NCBI

13 

Deng H, Yang F, Xu H, Sun Y, Xue X, Du S, Wang X, Li S, Liu Y and Wang R: Ac-SDKP suppresses epithelial-mesenchymal transition in A549 cells via HSP27 signaling. Exp Mol Pathol. 97:176–183. 2014. View Article : Google Scholar : PubMed/NCBI

14 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

15 

Harigai T, Kondo M, Isozaki M, Kasukawa H, Hagiwara H, Uchiyama H and Kimura J: Preferential binding of polyethylene glycol-coated liposomes containing a novel cationic lipid, TRX-20, to human subendthelial cells via chondroitin sulfate. Pharm Res. 18:1284–1290. 2001. View Article : Google Scholar : PubMed/NCBI

16 

Negishi Y, Endo-Takahashi Y, Matsuki Y, Kato Y, Takagi N, Suzuki R, Maruyama K and Aramaki Y: Systemic delivery systems of angiogenic gene by novel bubble liposomes containing cationic lipid and ultrasound exposure. Mol Pharm. 9:1834–1840. 2012. View Article : Google Scholar : PubMed/NCBI

17 

Arisaka M, Takano K, Negishi Y, Arima H and Aramaki Y: Involvement of lipid rafts in macrophage apoptosis induced by cationic liposomes. Arch Biochem Biophys. 508:72–77. 2011. View Article : Google Scholar : PubMed/NCBI

18 

Bailey AL and Cullis PR: Membrane fusion with cationic liposomes: effects of target membrane lipid composition. Biochemistry. 36:1628–1634. 1997. View Article : Google Scholar : PubMed/NCBI

19 

Bajoria R, Sooranna S and Chatterjee R: Effect of lipid composition of cationic SUV liposomes on materno-fetal transfer of warfarin across the perfused human term placenta. Placenta. 34:1216–1222. 2013. View Article : Google Scholar : PubMed/NCBI

20 

Barenholz Y, Bombelli C, Bonicelli MG, di Profio P, Giansanti L, Mancini G and Pascale F: Influence of lipid composition on the thermotropic behavior and size distribution of mixed cationic liposomes. J Colloid Interface Sci. 356:46–53. 2011. View Article : Google Scholar : PubMed/NCBI

21 

Bianco A, Napolitano R, Bonadies F, Celona D, Ortaggi G and Cametti C: Liposomes from a new chiral cationic lipid based on iridoidic template. Natl Prod Res. 21:1221–1227. 2007. View Article : Google Scholar

22 

Chen T, Wang RT, Wang Z, Lu TL and Zhao W: Construction and evaluation of non-specific targeting cationic polymer lipid liposomes. Yao Xue Xue Bao. 45:359–364. 2010.PubMed/NCBI

23 

de Paula Rigoletto T, Silva CL, Santana MH, Rosada RS and de la Torre LG: Effects of extrusion, lipid concentration and purity on physico-chemical and biological properties of cationic liposomes for gene vaccine applications. J Microencapsul. 29:759–769. 2012. View Article : Google Scholar : PubMed/NCBI

24 

Wang T, Zhen Y, Ma X, Wei B, Li S and Wang N: Mannosylated and lipid A-incorporating cationic liposomes constituting microneedle arrays as an effective oral mucosal HBV vaccine applicable in the controlled temperature chain. Colloids Surf B Biointerfaces. 126:520–530. 2015. View Article : Google Scholar : PubMed/NCBI

25 

Wang N, Wang T, Zhang M, Chen R, Niu R and Deng Y: Mannose derivative and lipid A dually decorated cationic liposomes as an effective cold chain free oral mucosal vaccine adjuvant-delivery system. Eur J Biopharm. 88:194–206. 2014. View Article : Google Scholar

26 

Vidyasagar A, Wilson NA and Djamali A: Heat shock protein 27 (HSP27): Biomarker of disease and therapeutic target. Fibrogenesis Tissue Repair. 5:72012. View Article : Google Scholar : PubMed/NCBI

27 

Zhu Y, Liu Y, Qian Y, Dai X, Yang L, Chen J, Guo S and Hisamitsu T: Research on the efficacy of Celastrus Orbiculatus in suppressing TGF-β1-induced epithelial-mesenchymal transition by inhibiting HSP27 and TNF-α-induced NF-κB/Snail signaling pathway in human gastric adenocarcinoma. BMC Complement Altern Med. 14:4332014. View Article : Google Scholar : PubMed/NCBI

28 

Cordonnier T, Bishop JL, Shiota M, Nip KM, Thaper D, Vahid S, Heroux D, Gleave M and Zoubeidi A: Hsp27 regulates EGF/β-catenin mediated epithelial to mesenchymal transition in prostate cancer. Int J Cancer. 136:E496–E507. 2015. View Article : Google Scholar : PubMed/NCBI

29 

Chen SF, Nieh S, Jao SW, Liu CL, Wu CH, Chang YC, Yang CY and Lin YS: Quercetin suppresses drug-resistant spheres via the p38 MAPK-Hsp27 apoptotic pathway in oral cancer cells. PLoS One. 7:e492752012. View Article : Google Scholar : PubMed/NCBI

30 

Mizutani H, Okano T, Minegishi Y, Matsuda K, Sudoh J, Kitamura K, Noro R, Soeno C, Yoshimura A, Seike M and Gemma A: HSP27 modulates epithelial to mesenchymal transition of lung cancer cells in a Smad-independent manner. Oncol Lett. 1:1011–1016. 2010. View Article : Google Scholar : PubMed/NCBI

31 

Shiota M, Bishop JL, Nip KM, Zardan A, Takeuchi A, Cordonnier T, Beraldi E, Bazov J, Fazli L, Chi K, et al: Hsp27 regulates epithelial mesenchymal transition, metastasis and circulating tumor cells in prostate cancer. Cancer Res. 73:3109–3119. 2013. View Article : Google Scholar : PubMed/NCBI

32 

Wei L, Liu TT, Wang HH, Hong HM, Yu AL, Feng HP and Chang WW: Hsp27 participates in the maintenance of breast cancer stem cells through regulation of epithelial-mesenchymal transition and nuclear factor-κB. Breast Cancer Res. 13:R1012011. View Article : Google Scholar : PubMed/NCBI

33 

Bernard Y, Ribeiro N, Thuaud F, Türkeri G, Dirr R, Boulberdaa M, Nebigil CG and Désaubry L: Flavaglines alleviate doxorubicin cardiotoxicity: Implication of Hsp27. PLoS One. 6:e253022011. View Article : Google Scholar : PubMed/NCBI

34 

Won YW, Kim JK, Cha MJ, Hwang KC, Choi D and Kim YH: Prolongation and enhancement of the anti-apoptotic effects of PTD-Hsp27 fusion proteins using an injectable thermo-reversible gel in a rat myocardial infarction model. J Control Release. 144:181–189. 2010. View Article : Google Scholar : PubMed/NCBI

35 

Shen Y, Miao N, Xu J, Gan X, Xu D, Zhou L, Xue H, Zhang W and Lu L: Metformin prevents renal fibrosis in mice with unilateral ureteral obstruction and inhibits ang II-induced ECM production in renal fibroblasts. Int J Mol Sci. 17:2016. View Article : Google Scholar

36 

Minton K: Extracellular matrix: Preconditioning the ECM for fibrosis. Nat Rev Mol Cell Biol. 15:766–767. 2014. View Article : Google Scholar : PubMed/NCBI

37 

Li SB and Jia YJ: Interactions between ECM and HSC cells in hepatic fibrosis. Sheng Li Ke Xue Jin Zhan. 45:462–464. 2014.(In Chinese). PubMed/NCBI

38 

Corona BT, Wu X, Ward CL, McDaniel JS, Rathbone CR and Walters TJ: The promotion of a functional fibrosis in skeletal muscle with volumetric muscle loss injury following the transplantation of muscle-ECM. Biomaterials. 34:3324–3335. 2013. View Article : Google Scholar : PubMed/NCBI

39 

Chun TH: Peri-adipocyte ECM remodeling in obesity and adipose tissue fibrosis. Adipocyte. 1:89–95. 2012. View Article : Google Scholar : PubMed/NCBI

40 

Choi MC, Cheung KK, Li X and Cheing GL: Pulsed electromagnetic field (PEMF) promotes collagen fibre deposition associated with increased myofibroblast population in the early healing phase of diabetic wound. Arch Dermatol Res. 308:21–29. 2016. View Article : Google Scholar : PubMed/NCBI

41 

Al-Qattan MM, Abd-Elwahed MM, Hawary K, Arafah MM and Shier MK: Myofibroblast expression in skin wounds is enhanced by collagen III suppression. Biomed Res Int. 2015:9586952015. View Article : Google Scholar : PubMed/NCBI

42 

Jiang HS, Zhu LL, Zhang Z, Chen H, Chen Y and Dai YT: Estradiol attenuates the TGF-β1-induced conversion of primary TAFs into myofibroblasts and inhibits collagen production and myofibroblast contraction by modulating the Smad and Rho/ROCK signaling pathways. Int J Mol Med. 36:801–807. 2015. View Article : Google Scholar : PubMed/NCBI

43 

Mia MM and Bank RA: The pro-fibrotic properties of transforming growth factor on human fibroblasts are counteracted by caffeic acid by inhibiting myofibroblast formation and collagen synthesis. Cell Tissue Res. 363:775–789. 2016. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

May 2018
Volume 17 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Deng, H., Gao, X., Peng, H., Wang, J., Hou, X., Xu, H., & Yang, F. (2018). Effect of liposome‑mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells. Molecular Medicine Reports, 17, 7319-7324. https://doi.org/10.3892/mmr.2018.8744
MLA
Deng, H., Gao, X., Peng, H., Wang, J., Hou, X., Xu, H., Yang, F."Effect of liposome‑mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells". Molecular Medicine Reports 17.5 (2018): 7319-7324.
Chicago
Deng, H., Gao, X., Peng, H., Wang, J., Hou, X., Xu, H., Yang, F."Effect of liposome‑mediated HSP27 transfection on collagen synthesis in alveolar type II epithelial cells". Molecular Medicine Reports 17, no. 5 (2018): 7319-7324. https://doi.org/10.3892/mmr.2018.8744