Effects of RNA interference targeting STAT3 gene on proliferation and apoptosis of HeLa cells

【Objective】To construct eukaryotic vector expressing short hairpin RNA(shRNA) of signal transducers and activators of transcription 3(STAT3),and to observe its effects on proliferation and apoptosis of HeLa cells.【Methods】shRNA templates was designed based on STAT3 gene sequence and was cloned into pSilencer2.1-U6-neo vector.The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000.The STAT3 protein and mRNA were detected by Western blotting and RT-PCR,respectively.The cellular growth activity was assayed by MTT,and the apoptosis was tested by flow cytometry.【Results】The plasmid pSilencer2.1-U6-neo-STAT3 was successfully constructed and transfected into HeLa cells.The protein of STAT3,the mRNA of STAT3,the ratio of cellular growth activity,and the ratio of cell apoptosis in nonspecific siRNA group were 0.55±0.08,0.49±0.05,(4.12±0.45)%,and(11.31±1.69)%,The protein of STAT3,the mRNA of STAT3,the ratio of cellular growth activity,and the ratio of cell apoptosis in target siRNA group were 0.36±0.06,0.32±0.05,(2.51±0.48)%,and(17.39±1.58)%.Contrasted with nonspecific siRNA group,the expressions of STAT3 in target siRNA group decreased,The cellular growth activity decreased,and cell apoptosis increased.【Conclusions】The siRNA expressing plasmid pSilencer2.1-U6-neo-STAT3 could inhibit the cellular proliferation and induce apoptosis of cervical cancer cells through suppressing the expression of STAT3.