Abstract:
Objective To investigate the effect of the short hairpin RNA ( shRNA) against human telomerase reverse t ranscriptase (hTERT) on the proliferation and apoptosis of prostate cancer cells PC-3m in vitro. Methods The recombinant plasmid psilencer-TRT was transfected into prostate cancer cell line PC-3m via liposome reagent . The level of hTERT mRNA was examined by reverse t ranscription polymerase chain reaction (RT-PCR) . The expressions of hTERT and c-myc protein were detected by western blot analysis. The effect of hTERT shRNA on the cellular proliferation capacity of PC-3m cells was assayed by the growth curve. The cell apoptosis was detected by Hoechst33258 staining, elect ron microscope, and flow cytomet ry analysis. Results The vector-mediated shRNA significantly reduced the level of hTERT mRNA by 89. 02 % af ter psilencer-TRT transduction in PC-3m cells. Meanwhile, the levels of hTERT and c-myc protein were also decreased in transfected cells. The cell proliferation was markedly inhibited compared with the cont rol cells. Partial cancer cells presented morphological changes of apoptosis, and the apoptosis rate was (19. 69 ±4. 75) %. Conclusion hTERT shRNA can suppress hTERT expression and cell proliferation, in addition to acceleration of apoptosis. This implied the possibility of RNA interfering to hTERT as the potential method for gene therapy of prostate cancer.