TERT promoter regulating melittin expression induces apoptosis and G<sub>0</sub>/G<sub>1</sub> cell cycle arrest in esophageal carcinoma cells

Zhou,Chao; Ma,Jie; Lu,Yuanhua; Zhao,Wan; Xu,Bingxue; Lin,Jian; Ma,Yongjun; Tian,Yafei; Zhang,Qi; Wang,Wei; Yan,Weiqun; Jiao,Ping

doi:10.3892/ol.2020.12277

January-2021 Volume 21 Issue 1

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January-2021 Volume 21 Issue 1

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Article Open Access

TERT promoter regulating melittin expression induces apoptosis and G₀/G₁ cell cycle arrest in esophageal carcinoma cells

Authors:
- Chao Zhou
- Jie Ma
- Yuanhua Lu
- Wan Zhao
- Bingxue Xu
- Jian Lin
- Yongjun Ma
- Yafei Tian
- Qi Zhang
- Wei Wang
- Weiqun Yan
- Ping Jiao
View Affiliations / Copyright

Affiliations: Department of Regenerative Medicine, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China

Copyright: © Zhou et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Article Number: 16
|
Published online on: November 6, 2020

https://doi.org/10.3892/ol.2020.12277
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Abstract

Esophageal squamous cell carcinoma accounts for a large proportion of cancer-associated mortalities in both men and women. Melittin is the major active component of bee venom, which has been reported to possess anti-inflammatory, antibacterial and anti-cancer properties. The aim of the present study was to construct a tumor targeted recombinant plasmid [pc-telomerase reverse transcriptase (TERT)-melittin] containing a human TERT promoter followed by a melittin coding sequence and to explore the effects of this plasmid in esophageal cell carcinoma and investigate preliminarily the underlying mechanisms of this effect. TE1 cells were transfected with pcTERT-melittin and the resulting apoptosis was subsequently examined. The viability of TE1 cells transfected with pcTERT-melittin was measured using a Cell Counting Kit-8 assay, which indicated inhibited proliferation. The disruption of mitochondrial membranes and the concomitant production of reactive oxygen species demonstrated an inducible apoptotic effect of melittin in TE1 cells. Apoptotic cells were also counted using an Annexin V-FITC and PI double-staining assay. The upregulation of cleaved caspase-9, cleaved caspase-3, Bax and poly(ADP-ribose) polymerase 1 in pcTERT-melittin transfected TE1 cells, suggested that pcTERT-melittin-induced apoptosis was associated with the mitochondrial pathway. TE1 cells were also arrested in the G₀/G₁ phase when transfected with pcTERT-melittin, followed by the decline of CDK4, CDK6 and cyclin D1 expression levels. As cell invasion and metastasis are common in patients with esophageal cancer, a cell migration assay was conducted and it was found that pcTERT-melittin transfection reduced the migratory and invasive abilities of TE1 cells. The findings of the present study demonstrated that pcTERT-melittin may induce apoptosis of esophageal carcinoma cells and inhibit tumor metastasis.

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1	Pakzad R, Mohammadian-Hafshejani A, Khosravi B, Soltani S, Pakzed I, Mohammadian M, Salehiniya H and Momenimovahed Z: The incidence and mortality of esophageal cancer and their relationship to development in Asia. Ann Transl Med. 4:292016.PubMed/NCBI
2	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
3	Abnet CC, Arnold M and Wei WQ: Epidemiology of esophageal squamous cell carcinoma. Gastroenterology. 154:360–373. 2018. View Article : Google Scholar : PubMed/NCBI
4	Liu X, Zhang M, Ying S, Zhang C, Lin R, Zheng J, Zhang G, Tian D, Guo Y, Du C, et al: Genetic alterations in esophageal tissues from squamous dysplasia to carcinoma. Gastroenterology. 153:166–177. 2017. View Article : Google Scholar : PubMed/NCBI
5	Testa U, Castelli G and Pelosi E: Esophageal cancer: Genomic and molecular characterization, stem cell compartment and clonal evolution. Medicines (Basel). 4:672017. View Article : Google Scholar
6	Chen M, Shen M, Lin Y, Liu P, Liu X, Li X, Li A, Yang R, Ni W, Zhou X, et al: Adjuvant chemotherapy does not benefit patients with esophageal squamous cell carcinoma treated with definitive chemoradiotherapy. Radiat Oncol. 13:1502018. View Article : Google Scholar : PubMed/NCBI
7	Wu SX, Wang LH, Luo HL, Xie CY, Zhang XB, Hu W, Zheng AP, Li DJ, Zhang HY, Xie CH, et al: Randomised phase III trial of concurrent chemoradiotherapy with extended nodal irradiation and erlotinib in patients with inoperable oesophageal squamous cell cancer. Eur J Cancer. 93:99–107. 2018. View Article : Google Scholar : PubMed/NCBI
8	Amer MH: Gene therapy for cancer: Present status and future perspective. Mol Cell Ther. 2:272014. View Article : Google Scholar : PubMed/NCBI
9	Li J, Liang H, Liu J and Wang Z: Poly (amidoamine) (PAMAM) dendrimer mediated delivery of drug and pDNA/siRNA for cancer therapy. Int J Pharm. 546:215–225. 2018. View Article : Google Scholar : PubMed/NCBI
10	Kim HJ, Kim A, Miyata K and Kataoka K: Recent progress in development of siRNA delivery vehicles for cancer therapy. Adv Drug Deliv Rev. 104:61–77. 2016. View Article : Google Scholar : PubMed/NCBI
11	Zhou Z, Liu X, Zhu D, Wang Y, Zhang Z, Zhou X, Qiu N, Chen X and Shen Y: Nonviral cancer gene therapy: Delivery cascade and vector nanoproperty integration. Adv Drug Deliv Rev. 115:115–154. 2017. View Article : Google Scholar : PubMed/NCBI
12	Orsolic N: Bee venom in cancer therapy. Cancer Metastasis Rev. 31:173–194. 2012. View Article : Google Scholar : PubMed/NCBI
13	Rajasekaran G, Dinesh Kumar S, Nam J, Jeon D, Kim Y, Lee CW, Park IS and Shin SY: Antimicrobial and anti-inflammatory activities of chemokine CXCL14-derived antimicrobial peptide and its analogs. Biochim Biophys Acta Biomembr. 1861:256–267. 2019. View Article : Google Scholar : PubMed/NCBI
14	Sobral F, Sampaio A, Falcão S, Queiroz MJ, Calhelha RC, Vilas-Boas M and Ferreira IC: Chemical characterization, antioxidant, anti-inflammatory and cytotoxic properties of bee venom collected in Northeast Portugal. Food Chem Toxicol. 94:172–177. 2016. View Article : Google Scholar : PubMed/NCBI
15	Lim HN, Baek SB and Jung HJ: Bee venom and its peptide component melittin suppress growth and migration of melanoma cells via inhibition of PI3K/AKT/mTOR and MAPK Pathways. Molecules. 24:9292019. View Article : Google Scholar
16	Rady I, Siddiqui IA, Rady M and Mukhtar H: Melittin, a major peptide component of bee venom, and its conjugates in cancer therapy. Cancer Lett. 402:16–31. 2017. View Article : Google Scholar : PubMed/NCBI
17	Gajski G and Garaj-Vrhovac V: Melittin: A lytic peptide with anticancer properties. Environ Toxicol Pharmacol. 36:697–705. 2013. View Article : Google Scholar : PubMed/NCBI
18	Leao R, Apolonio JD, Lee D, Figueiredo A, Tabori U and Castelo-Branco P: Mechanisms of human telomerase reverse transcriptase (hTERT) regulation: Clinical impacts in cancer. J Biomed Sci. 25:222018. View Article : Google Scholar : PubMed/NCBI
19	Ly JD, Grubb DR and Lawen A: The mitochondrial membrane potential (deltapsi(m)) in apoptosis; an update. Apoptosis. 8:115–128. 2003. View Article : Google Scholar : PubMed/NCBI
20	Perelman A, Wachtel C, Cohen M, Haupt S, Shapiro H and Tzur A: JC-1: Alternative excitation wavelengths facilitate mitochondrial membrane potential cytometry. Cell Death Dis. 3:e4302012. View Article : Google Scholar : PubMed/NCBI
21	Rao XY, Huang XL, Zhou XC and Lin X: An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 3:71–85. 2013.PubMed/NCBI
22	Yang Y, Karakhanova S, Hartwig W, D'Haese JG, Philippov PP, Werner J and Bazhin AV: Mitochondria and mitochondrial ros in cancer: Novel targets for anticancer therapy. J Cell Physiol. 231:2570–2581. 2016. View Article : Google Scholar : PubMed/NCBI
23	Song SB, Jang SY, Kang HT, Wei B, Jeoun UW, Yoon GS and Hwang ES: Modulation of mitochondrial membrane potential and ROS generation by nicotinamide in a manner independent of SIRT1 and mitophagy. Mol Cells. 40:503–514. 2017.PubMed/NCBI
24	Lin M, Tang S, Zhang C, Chen H, Huang W, Liu Y and Zhang J: Euphorbia factor L2 induces apoptosis in A549 cells through the mitochondrial pathway. Acta Pharm Sin B. 7:59–64. 2017. View Article : Google Scholar : PubMed/NCBI
25	Wu R, Tang S, Wang M, Xu X, Yao C and Wang S: MicroRNA-497 induces apoptosis and suppresses proliferation via the Bcl-2/Bax-Caspase9-Caspase3 Pathway and Cyclin D2 Protein in HUVECs. PLoS One. 11:e01670522016. View Article : Google Scholar : PubMed/NCBI
26	Luch A: Cell cycle control and cell division: Implications for chemically induced carcinogenesis. Chembiochem. 3:506–516. 2002. View Article : Google Scholar : PubMed/NCBI
27	Otto T and Sicinski P: Cell cycle proteins as promising targets in cancer therapy. Nature reviews. Cancer. 17:93–115. 2017.PubMed/NCBI
28	Sherr CJ, Beach D and Shapiro GI: Targeting CDK4 and CDK6: From discovery to therapy. Cancer Discov. 6:353–367. 2016. View Article : Google Scholar : PubMed/NCBI
29	Chen J: The cell-cycle arrest and apoptotic functions of p53 in tumor initiation and progression. Cold Spring Harb Perspect Med. 6:a0261042016. View Article : Google Scholar : PubMed/NCBI
30	Lee G and Bae H: Anti-inflammatory applications of melittin, a major component of bee venom: Detailed mechanism of action and adverse effects. Molecules. 21:6162016. View Article : Google Scholar
31	Kong GM, Tao WH, Diao YL, Fang PH, Wang JJ, Bo P and Qian F: Melittin induces human gastric cancer cell apoptosis via activation of mitochondrial pathway. World J Gastroenterol. 22:3186–3195. 2016. View Article : Google Scholar : PubMed/NCBI
32	Soliman C, Eastwood S, Truong VK, Ramsland PA and Elbourne A: The membrane effects of melittin on gastric and colorectal cancer. PLoS One. 14:e02240282019. View Article : Google Scholar : PubMed/NCBI
33	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
34	Guo LD, Chen XJ, Hu YH, Yu ZJ, Wang D and Liu JZ: Curcumin inhibits proliferation and induces apoptosis of human colorectal cancer cells by activating the mitochondria apoptotic pathway. Phytother Res. 27:422–430. 2013. View Article : Google Scholar : PubMed/NCBI
35	Bishayee K, Ghosh S, Mukherjee A, Sadhukhan R, Mondal J and Khuda-Bukhsh AR: Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G₂/M, in cervical carcinoma: Signal cascade and drug-DNA interaction. Cell Prolif. 46:153–163. 2013. View Article : Google Scholar : PubMed/NCBI
36	Hu Q, Wu D, Chen W, Yan Z, Yan C, He T, Liang Q and Shi Y: Molecular determinants of caspase-9 activation by the Apaf-1 apoptosome. Proc Natl Acad Sci USA. 111:16254–16261. 2014. View Article : Google Scholar : PubMed/NCBI
37	Bai P: Biology of poly(ADP-Ribose) polymerases: The factotums of cell maintenance. Mol Cell. 58:947–958. 2015. View Article : Google Scholar : PubMed/NCBI
38	Los M, Mozoluk M, Ferrari D, Stepczynska A, Stroh C, Renz A, Herceg Z, Wang ZQ and Schulze-Osthoff K: Activation and caspase-mediated inhibition of PARP: A molecular switch between fibroblast necrosis and apoptosis in death receptor signaling. Mol Biol Cell. 13:978–988. 2002. View Article : Google Scholar : PubMed/NCBI
39	Jiang Y, Zhang J, Zhao J, Li Z, Chen H, Qiao Y, Chen X, Liu K and Dong Z: TOPK promotes metastasis of esophageal squamous cell carcinoma by activating the Src/GSK3β/STAT3 signaling pathway via γ-catenin. BMC Cancer. 19:12642019. View Article : Google Scholar : PubMed/NCBI
40	Giono LE and Manfredi JJ: The p53 tumor suppressor participates in multiple cell cycle checkpoints. J Cell Physiol. 209:13–20. 2006. View Article : Google Scholar : PubMed/NCBI
41	McCurrach ME, Connor TM, Knudson CM, Korsmeyer SJ and Lowe SW: Bax-deficiency promotes drug resistance and oncogenic transformation by attenuating p53-dependent apoptosis. Proc Natl Acad Sci USA. 94:2345–2349. 1997. View Article : Google Scholar : PubMed/NCBI

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