靶向STAT3基因的小干扰RNA对子宫内膜癌HEC-1B细胞生物学行为的影响

The effect of a small interfering RNA targeting STAT 3 gene on the biological function of human endometrial cancer cell line HEC-1B

目的 :探讨靶向信号转导与转录激活因子3(signal transducer and activator of transcription 3,STAT 3)基因的小干扰RNA(small interfering RNA,siRNA)对人子宫内膜癌HEC-1B细胞生物学行为的影响。方法 :将化学合成的靶向STAT 3基因的siRNA片段(si-STAT3)转染至HEC-1B细胞,实时荧光定量-PCR法检测转染后细胞中STAT3 mRNA的表达,CCK-8(cell counting kit-8)法检测细胞的增殖活性,FCM法检测细胞周期和细胞凋亡,Transwell小室法检测细胞的迁移和侵袭,蛋白质印迹法检测细胞中STAT3、磷酸化STAT3及其下游细胞周期蛋白D2(cyclin D2,CCND2)和基质金属蛋白酶2(metalloproteinase-2,MMP-2)蛋白的表达。结果 :与空白对照组(HEC-1B细胞中仅加入脂质体)或转染对照干扰片段(si-control)组比较,转染si-STAT3后的HEC-1B细胞中STAT3 mRNA的表达水平明显降低(P<0.01),细胞的增殖活性明显下降(P<0.01),G1期细胞所占比例上升,S期细胞所占比例下降(P<0.01),细胞早期凋亡率明显升高(P<0.01),细胞的迁移和侵袭能力明显下降(P<0.01),HEC-1B细胞中STAT3、磷酸化STAT3及其下游CCND2和MMP-2蛋白的表达水平明显下调(P<0.01)。结论 :靶向STAT 3基因的特异性si-RNA片段能够下调HEC-1B细胞中STAT3mRNA及其蛋白的表达,抑制HEC-1B细胞的增殖、侵袭和迁移,并诱导其凋亡。

Objective： To investigate the effect of a small interfering RNA （si-RNA） targeting signal transducer and activator of transcription-3 （STAT3） on the biological function of human endometrial cancer cell line HEC-1B. Methods： The HEC-1B cells were transfected with chemically synthesized si-RNA targeting STAT3 （si-STAT3）. Then the expression of STAT3 mRNA was detected by real-time fluorescence quantitative-PCR, the proliferation ability was determined by cell counting kit-8 （CCK-8） assay, the cell cycle distribution and apoptosis were measured by flow cytometry, the migration and invasion were examined by Transwell chamber, and the expression levels of STAT3, phospho-STAT3 （p-STAT3）, and cyclin D2 （CCND2） and metalloproteinase-2 （MMP-2） were detected by Western blotting. Results： As compared with the blank control group （only liposome was added） or transfection with si-controlgroup, the expression level of STAT3 mRNA in HEC-1B cells after transfection with si-STAT3 was down- regulated （P 〈 0.01）, the proliferation ability was inhibited （P 〈 0.01）, the percentage of the cells in G1- phase was increased and which in Sl-phase was decreased （P 〈 0.01）. The abilities of migration and invasion of HEC-1B cells after transfection with si-STAT3 were inhibited （P 〈 0.01）, and the expression levels of STAT3, p-STAT3, CCND2 and MMP-2 proteins were down-regulated （P 〈 0.01）. Conclusion： Si- RNA targeting STAT3 gene can inhibit the expressions of STAT3 mRNA and protein in HEC-1B cells, and also inhibit the cell proliferation, the abilities of migration and invasion, and apoptosis.