RNA Interference-mediated Significant Enhancement Of Apoptosis In Yunnan Gejiu Lung Squamous Carcinoma Cell YTMLC (Yunnan Tin Mineral Lung Carcinoma) And HeLa Cell Line By SiRNA Expression Vector Targeting Survivin Gene

Objective To apply the small interfering RNAs targeting survivin to inhibit expression of endogenous survivin gene in Yunnan Tin Mineral Lung Carcinoma(YTMLC) cell line and Hela cell line and to increase the apoptosis of YTMLC cell and Hela cell.Methods The recombinant plasmid pshRNA-survivin-1 and pshRNA-survivin-2 were constructed and transfected into YTMLC cell and Hela cell ,respectively. The expression level of survivin was determined by immunofluorescence staining and Western blot .The transcription of survivin gene was detected by semi-quantitative RT-PCR. The proliferation of YTMLC cell and Hela cell was detected by MTT method. The apoptosis in YTMLC cell and Hela cell were detected by PI-Annexin V staining.Results The introduction of plasmids pshRNA-survivin-1 and pshRNA-survivin-2 were showed to efficiently and specifically inhibit the expression of survivin according to results of immunofluorescence staining and Western blot. Semi-quantitative RT-PCR showed that mRNA transcription of survivin gene in YTMLC cell was reduced by nearly 80.60%and 70.09%, respectively. Semi-quantitative RT-PCR showed that mRNA transcription of survivin gene in Hela cell was reduced by nearly 81.04 %and 87.05%, respectively. On the contrast, the control plasmid did exhibit no inhibitory effect on the protein expression and mRNA transcription of survivin gene in YTMLC cell and Hela cell. The proliferation of YTMLC cell and Hela cell which were detected by MTT method both decrease distinctively. The apoptosis in YTMLC cell line were detected by PI-Annexin V staining with apoptosis rates at 33.42±2.42% (P<0.01) and 20.09±1.32% (P<0.01) , respectively. While,The apoptosis in Hela cell line were detected by PI-Annexin V staining with apoptosis rates at 36.02±2.12%(P<0.01 )and 35.29±2.02%(P<0.01), respectively.Conclusion The results demonstrate that the small interfering RNA targeting survivin gene shows dramastic inhibition on protein expression and RNA transcription and cell proliferation and significant enhancement of apoptosis in YTMLC cell line and Hela cell line, and should be a new strategy in gene silence therapy targeting survivin.