Effects of siRNA against Survivin gene on the proliferation and apoptosis of human renal carcinoma cell line 786-O
Kui-jun Wang, Bin Xu, Xi-guo Sun, Fangkun Zhou, Cunjie Li
Abstract
Kui-jun Wang, Bin Xu, Xi-guo Sun, Fangkun Zhou, Cunjie Li
Abstract
Objective To evaluate the effects of small interfering RNA (siRNA) against Survivin gene on the proliferation and apoptosis of human renal carcinoma cell line 786-O cells. Methods One pair of Survivin target sequence-specific siRNA was designed and synthesized, then siRNA/liposome complex was used to transfect the human renal carcinoma 786-O cells with increasing concentrations (10, 50, 100 nmol/L). The mRNA and protein expression of Survivin was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The proliferation of 786-O cells was measured by MTT assay. The apoptosis of 786-O cells was examined by TUNEL assay. Results SiRNA-Survivin efficiently down-regulated the Survivin expression, inhibited the proliferation and induced the apoptosis in a dose dependent manner (for mRNA 88. 3% , 62.4% , 43. 8% , for protein 87. 7% , 62.4% , 46.5%, proliferation-inhibiting rate 11.6%, 41.2%, 57.5%, and apoptosis rate 14.2%, 29.4%, 38. 1% , respectively) (P<0.05). Conclusion siRNA against Survivin gene can inhibit the proliferation, respectively) (P<0.05). Conclusion siRNA against Survivin gene can inhibit the proliferation Key words: Renal cell carcinoma; Survivin gene; Small interfering RNA
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Objective To evaluate the effects of small interfering RNA (siRNA) against Survivin gene on the proliferation and apoptosis of human renal carcinoma cell line 786-O cells. Methods One pair of Survivin target sequence-specific siRNA was designed and synthesized, then siRNA/liposome complex was used to transfect the human renal carcinoma 786-O cells with increasing concentrations (10, 50, 100 nmol/L). The mRNA and protein expression of Survivin was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. The proliferation of 786-O cells was measured by MTT assay. The apoptosis of 786-O cells was examined by TUNEL assay. Results SiRNA-Survivin efficiently down-regulated the Survivin expression, inhibited the proliferation and induced the apoptosis in a dose dependent manner (for mRNA 88. 3% , 62.4% , 43. 8% , for protein 87. 7% , 62.4% , 46.5%, proliferation-inhibiting rate 11.6%, 41.2%, 57.5%, and apoptosis rate 14.2%, 29.4%, 38. 1% , respectively) (P<0.05). Conclusion siRNA against Survivin gene can inhibit the proliferation, respectively) (P<0.05). Conclusion siRNA against Survivin gene can inhibit the proliferation Key words: Renal cell carcinoma; Survivin gene; Small interfering RNA
Key concepts: Survivin, Small interfering RNA, Transfection, Apoptosis, Molecular biology, Cell growth, Cancer research, Cell culture