JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2015, Vol. 53 ›› Issue (1): 27-33.doi: 10.6040/j.issn.1671-7554.0.2014.576

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Inhibitive effects of lenti virus-mediated hTERT RNA interference and TMPyP4-PDT on cervix cancer SiHa cells

WANG Jie, ZHANG Youzhong, FENG Jinbo, LIU Hongli, ZONG Liju   

  1. Department of Genecology and Obstetrics, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China
  • Received:2014-09-03 Revised:2014-10-29 Published:2015-01-10

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Abstract: Objective To investigate the effects of lenti-hTERT-shRNA combined with photodynamic therapy mediated by meso-5,10,15,20-Tetrakis-(Nmethyl-4-pyridyl) (TMPyP4) on cervical cancer SiHa cells in vitro. Methods Four groups were enrolled in the experiment, including the blank control group, gene therapy group, PDT group and combined treatment group. Lenti-hTERT-shRNA was synthesized and SiHa cells were transfected, then the optimum multiplicity of infection (MOI) was confirmed. After that, the cells were treated with 7.5 μmol/L TMPyP4 and irradiation (blue laser, 4 J/cm2). The proliferative activity of SiHa cells was assessed by Cell Counting Kit (CCK-8) assay; the expressions of hTERT, p53, HPV-16 E6, and Caspase-8 mRNA were detected by real-time PCR, and the hTERT protein was tested by immunofluorescence (IF); the apoptosis was examined by AnnexinV-PE/7-AAD double dye kit; the ability of cellular invasion was assessed by Transwell experiment. Results Compared with the blank control group, the other three groups had decreased the expressions of hTERT, HPV-16 E6, and Caspase-8 mRNA, but increased the expression of p53 mRNA. The down-regulation of hTERT protein was the most significant in the combined treatment group (P<0.05). In the combined treatment group, the cellular survivability and invasion ability of SiHa cells were inhibited more markedly than in the other groups (P<0.05), while the apoptosis rate increased more significantly (P<0.05). Conclusion Lenti virus-mediated hTERT RNA interference combined with photodynamic therapy of TMPyP4can suppress the abilities of proliferation and invasion, and enhance the apoptosis rate of cervix cancer SiHa cells.

Key words: Photodynamic therapy, SiHa cells, Cervix neoplasms, Human telomerase reverse transcriptase, Meso-5,10,15, 20-Tetrakis- (Nmethyl-4-pyridyl) porphine

CLC Number: 

  • R454.2
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