Effect of hTERT Gene Expression on Proliferation and Apoptosis of Cervical Cancer Caski Cells by Lentivirus-Mediated Small Interfering RNA

doi:10.11910/2227-6394.2016.04.02.07

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Abstract

Objective: To explore the feasibility of lentivirus-mediated small interfering RNA (siRNA) silencing the human telomerase reverse transcriptase (hTERT) and its influence on the proliferation and apoptosis of cervical cancer Caski cells.
Methods: The cervical cancer Caski cells were transfected by lentivirus expression vectors anti-hTERT-TV and NC-LV which had been constructed in advance. There were three groups in the experiment, namely blank group (NC group), negative control group (NC-LV group) and interference group (anti-hTERT-LV group). The expression level of hTERT mRNA was determined by real-time fluorescent quantitative polymerase chain reaction (PCR), while the cell cycle and apoptosis were detected by flow cytometry.
Results: The relative expression level of hTERT mRNA in anti-hTERT-LV group was significantly lower than those in NC group and NC-LV group (P<0.001). The proportion of cells in G1 phase in anti-hTERT-LV group was significantly higher than those in NC group and NC-LV group (P<0.001), while that in S phase was significantly lower than those in NC group and NC-LV group (P<0.001). The number of cell clones in anti-hTERT-LV group was significantly lower than those in NC group and NC-LV group (P<0.001), but there was no statistical significance between NC-LV group and NC group (P>0.05).
Conclusion: The lentivirus-mediated siRNA can induce silencing hTERT gene, effectively suppress its expression, promote the cell apoptosis and inhibit the proliferation of cervical cancer Caski cells.

Key words： Cervical cancer Caski cells hTERT Cell proliferation Apoptosis Cell cycle

Fund: This study was approved by Guangxi Natural Science Foundation Project (No.2013GXNSFAA019254).

Corresponding Authors: LING Dan, E-mail: 1279512058@qq.com

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	MO Ling-zhao
	LI De-quan
	XIA Yu
	CHEN Chang-xian
	PANG Xiao-fen
	LING Dan

Cite this article:

MO Ling-zhao,LI De-quan,XIA Yu, et al. Effect of hTERT Gene Expression on Proliferation and Apoptosis of Cervical Cancer Caski Cells by Lentivirus-Mediated Small Interfering RNA[J]. Journal of International Translational Medicine, 2016, 4(2): 114-120.

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http://www.jitm.hk/EN/10.11910/2227-6394.2016.04.02.07 OR http://www.jitm.hk/EN/Y2016/V4/I2/114

1. Song YJ, Kim JY, Lee SK, et al. Persistent human papillomavirus DNA is associated with local recurrence after radiotherapy of uterine cervical cancer. Int J Cancer, 2011, 129(4): 896-902. doi:10.1002/ijc.25741.
2. Kirkegård J, Farkas DK, Søgaard M, et al. Conization as a marker of persistent cervical human papillomavirus (HPV) infection and risk of gastrointestinalcancer: a Danish 34-year nationwide cohort study. Cancer Causes Control, 2014, 25(12): 1677-1682. doi: 10.1007/s10552-014-0473-4.
3. Katki HA, Cheung LC, Fetterman B, et al. A joint model of persistent human papillomavirus infection and cervical cancer risk: Implications for cervical cancer screening. J R Stat Soc Ser A Stat Soc, 2015, 178(4): 903-923. doi: 10.1111/rssa.12101.
4. Zhou C, Steplowski TA, Dickens HK, et al. Estrogen induction of telomerase activity through regulation of the mitogen-activated protein kinase (MAPK) dependent pathway in human endometrial cancer cells. PLoS One, 2013, 8(2): e55730. doi: 10.1371/journal.pone.0055730.
5. Miller J, Dakic A, Chen R, et al. HPV16 E7 protein and hTERT proteins defective for telomere maintenance cooperate to immortalize human keratinocytes. PLoS Pathog, 2013, 9(4): e1003284. doi: 10.1371/journal.ppat.1003284.
6. Peralta-Zaragoza O, De-la-O-Gómez F, Deas J, et al. Selective silencing of gene target expression by siRNA expression plasmids in human cervical cancer cells. Methods Mol Biol, 2015, 1249: 153-171. doi: 10.1007/978-1-4939-2013-6_11.
7. Díaz-González Sdel M, Deas J, Benítez-Boijseauneau O, et al. Utility of microRNAs and siRNAs in cervical carcinogenesis. Biomed Res Int, 2015, 2015: 374924. doi: 10.1155/2015/374924.
8. Shi YA, Zhao Q, Zhang LH, et al. Knockdown of hTERT by siRNA inhibits cervical cancer cell growth in vitro and in vivo. Int J Oncol, 2014, 45(3): 1216-1224. doi: 10.3892/ijo.2014.2493.
9. Nachajova M, Brany D, Dvorska D. Telomerase and the process of cervical carcinogenesis. Tumour Biol, 2015, 36(10): 7335-7338. doi: 10.1007/s13277-015-3976-z.
10. Sui X, Kong N, Wang Z, et al. Epigenetic regulation of the human telomerase reverse transciptase gene: A potential therapeutic target for the treatment of leukemia. Oncol Lett, 2013, 6(2): 317-322. doi: 10.3892/ol.2013.1367.
11. Merghoub N, El Btaouri H, Benbacer L, et al. Inula viscosa extracts induces telomere shortening and apoptosis in cancer cells and overcome drug resistance. Nutr Cancer, 2016, 68(1): 131-143. doi: 10.1080/01635581.2016.1115105.
12. Iarriccio L, Manguán-García C, Pintado-Berninches L, et al. GSE4, a small dyskerin- and GSE24.2-related peptide, induces telomerase activity, cell proliferation and reduces DNA damage, oxidative stress and cell senescence in dyskerin mutant cells. PLoS One, 2015, 10(11): e0142980. doi: 10.1371/journal.pone.0142980.
13. Hahn WC, Stewart SA, Brooks MW, et al. Inhibition of telomerase limits the growth of human cancer cells. Nat Med, 1999, 5(10): 1164-1170. doi: 10.1038/13495.
14. Panneer Selvam S, De Palma RM, Oaks JJ, et al. Binding of the sphingolipid S1P to hTERT stabilizes telomerase at the nuclear periphery by allosterically mimicking protein phosphorylation. Sci Signal, 2015, 8(381): ra58. doi: 10.1126/scisignal.aaa4998.
15. Gottumukkala SN, Dwarakanath CD, Sudarsan S．Ribonucleic acid interference induced gene knockdown. J Indian Soc Periodontol, 2013, 17(4): 417-422. doi: 10.4103/0972-124X.118309.
16. Ranjbar B, Krogh LB, Laursen MB, et al. Anti-apoptotic effects of lentiviral vector transduction promote increased rituximab tolerance in cancerous B-cells. PLoS One, 2016, 11(4): e0153069. doi: 10.1371/journal.pone.0153069.
17. Dreyer JL. Lentiviral vector-mediated gene transfer and RNA silencing technology in neuronal dysfunctions. Mol Biotechnol, 2011, 47(2): 169-187. doi: 10.1007/s12033-010-9334-x.
18. Zhao HK, Mo LZ. Construction of the lentiviral expression vector of human telomerase reverse transcriptase gene small interfering RNA fragments and its expression in cervical cancer caski cells. Chinese J Cancer Prev Contr, 2014, 6(4): 342-347.