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目的探讨LIM同源盒基因8(Lhx8)对卵巢癌细胞株SKOV3增殖、转移和侵袭的影响。方法采用Lhx8过表达慢病毒(LV-Lhx8)转染卵巢癌细胞株SKOV3构建过表达模型;设阴性对照慢病毒(LV-NC)组通过Lipofectamine 2000转染Lhx8-siRNA构建细胞干扰模型并设阴性对照序列(FAM-siRNA;NC)组;野生(WT)组细胞仅给予等量不含病毒或任何干扰片段的转染试剂。通过免疫荧光、蛋白质印迹法和qPCR验证过表达和干扰效果。采用EDU和细胞周期实验检测过表达或干扰Lhx8后细胞的增殖能力;采用Transwell和划痕实验分别检测转染后细胞的迁移和侵袭能力。结果 Lhx8过表达组SKOV3细胞中Lhx8的表达高于LV-NC和WT组(P<0.01),干扰后Lhx8的mRNA和蛋白表达均降低,与NC和WT组相比差异有统计学意义(P<0.01)。与WT和LV-NC组相比,Lhx8过表达抑制SKOV3细胞的增殖(P<0.01),而干扰Lhx8后其增殖能力增加(P<0.01);细胞周期实验结果示过表达Lhx8能够增加进入G0/G1期的细胞数目,从而抑制细胞增殖(P<0.01),干扰Lhx8表达后进入S期的细胞数目多于WT和NC组(P<0.01)。划痕和Transwell实验结果示SKOV3细胞过表达Lhx8后其迁移和侵袭能力均低于WT和LV-NC组(P<0.01),干扰Lhx8表达后细胞的迁移和侵袭能力均高于WT和NC组(P<0.01)。SKOV3细胞过表达Lhx8后基质金属蛋白酶(MMP)-2和MMP-9的表达均低于WT和LV-NC组(P<0.01),干扰Lhx8后MMP-2、MMP-9表达均高于WT和NC组(P<0.01)。结论 Lhx8能够抑制SKOV3细胞的增殖、迁移和侵袭,同时能够下调MMP-2和MMP-9的表达。
Objective To investigate the effect of LIM homeobox gene 8 (Lhx8) on the proliferation, metastasis and invasion of ovarian cancer cell line SKOV3. Methods Ovarian cancer cell line SKOV3 was transfected with Lhx8 overexpression lentivirus (LV-Lhx8) to construct an over-expression model. The negative control lentivirus (LV-NC) group was transfected with Lhx8-siRNA by Lipofectamine 2000, Control group (FAM-siRNA; NC) group; wild-type (WT) group cells were given only the same amount of transfection reagent containing no virus or any interfering fragments. Overexpression and interference effects were verified by immunofluorescence, Western blotting and qPCR. Cell proliferation was assessed by EDU and cell cycle assay after Lhx8 overexpression or interference. Transwell and scratch assays were used to detect the migration and invasion ability of transfected cells. Results The expression of Lhx8 in LOVx overexpression group was higher than that in LV-NC and WT groups (P <0.01), and the mRNA and protein expression of Lhx8 in LOVx overexpression group was lower than that in NC and WT groups (P <0.01). Compared with WT and LV-NC groups, Lhx8 overexpression inhibited the proliferation of SKOV3 cells (P <0.01), but increased the proliferation ability of Lhx8 cells after interference with Lhx8 (P <0.01). The results of cell cycle showed that Lhx8 overexpression increased the expression of G0 / G1 phase cells (P <0.01). The number of cells that interfered with Lhx8 expression in S phase was more than that in WT and NC groups (P <0.01). Scratches and Transwell results showed that the migration and invasion ability of SKOV3 cells after Lhx8 overexpression was lower than that of WT and LV-NC groups (P <0.01), and the migration and invasion ability of LOVx cells were higher than that of WT and NC groups (P <0.01). The expression of MMP-2 and MMP-9 in SKOV3 cells after Lhx8 overexpression was lower than that in WT and LV-NC groups (P <0.01) And NC group (P <0.01). Conclusion Lhx8 can inhibit the proliferation, migration and invasion of SKOV3 cells and down-regulate the expressions of MMP-2 and MMP-9.