沉默PTX3表达对子宫内膜癌细胞生物学行为的影响

Effect of silencing PTX3 expression on biological behavior of endometrial cancer cells

目的探讨沉默正五聚蛋白3(PTX3)表达对子宫内膜癌细胞生物学行为的影响。方法采用qRT-PCR法和Western blotting法检测子宫内膜癌JEC、HEC-1B、KLE、Ishikawa细胞及子宫内膜上皮HEECs细胞PTX3 mRNA和蛋白表达,筛选PTX3表达最高的子宫内膜癌细胞进行后续实验。取传3代、对数生长期子宫内膜癌细胞,随机分为siRNA-PTX3组和siRNA-NC组,按Lipofectamine^(TM)2000说明分别转染siRNA-PTX3、siRNA-NC。收集两组转染24 h细胞,采用CCK-8法检测再培养24、48、72、96 h细胞增殖活性,采用Annexin V-FITC/PI双染法检测细胞凋亡率,采用流式细胞术检测细胞周期分布,采用Transwell小室实验检测细胞侵袭和迁移能力。结果各子宫内膜癌细胞PTX mRNA和蛋白相对表达量均明显高于子宫内膜上皮HEECs细胞(P均<0.05),且以Ishikawa细胞PTX mRNA和蛋白相对表达量最高。siRNA-PTX3组再培养48、72、96 h细胞增殖活性均低于siRNA-NC组同期(P均<0.05);与siRNA-NC组比较,siRNA-PTX3组细胞凋亡率明显升高,并且其G_0/G_1期细胞比例明显升高,S期细胞比例明显降低(P<0.05或<0.01)。siRNA-PTX3组细胞侵袭和迁移能力均明显低于siRNA-NC组(P均<0.01)。结论沉默PTX3表达能够抑制子宫内膜癌细胞增殖、侵袭和迁移,并诱导其凋亡。

Objective To investigate the effect of silencing pentraxin-3(PTX3)expression on the biological behavior of endometrial cancer cells.Methods The expression of PTX3 in JEC,HEC-1 B,KLE,and Ishikawa cells was detected by qRT-PCR and Western blotting.Ishikawa cells with the highest expression of PTX3 were screened out for subsequent experiments.Ishikawa cells in logarithmic growth phase of the third generation were randomly divided into the siRNA-PTX3 group and siRNA-NC group.According to the instructions of LipofectamineTM2000,siRNA-PTX3 and siRNA-NC were transfected into the cells of each group.Cells of the two groups were collected at 24 h after transfection.CCK-8 was used to detect the cell proliferation,Annexin V-FITC/PI double staining method was used to detect the apoptotic rate,flow cytometry was used to detect cell cycle distribution,and Transwell chamber test was used to detect cell invasion and migration abilities.Results The cell proliferation activity in the siRNA-PTX3 group was lower than that in the siRNA-NC group at 48,72 and 96 h(all P<0.05).Compared with the siRNA-NC group,the apoptotic rate of the siRNA-PTX3 group increased,and the proportion of cells in the G0/G1 phase increased,while the proportion of cells in the S phase decreased(all P<0.05 or all P<0.01).The invasion and migration abilities of cells in the siRNA-PTX3 group were significantly lower than those in the siRNA-NC group(both P<0.05).Conclusion Silencing PTX3 expression can inhibit the proliferation,invasion,and migration of endometrial cancer cells and induce the apoptosis.