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| LIM同源盒基因8对卵巢癌细胞SKOV3生物学行为的影响 |
| 黄铮1,马伟1,陆欢1,王斌1,金国华2,吕广明2,张新化2,张睿3* |
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(1. 上海交通大学附属第六人民医院南院妇产科, 上海 201499;
2. 南通大学医学院解剖教研室, 南通 226001;
3. 上海交通大学附属第六人民医院妇产科, 上海 200233
*通信作者) |
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| 摘要: |
| 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。 |
| 关键词: 卵巢肿瘤 LIM同源盒基因8 细胞增殖 细胞迁移 肿瘤侵袭 |
| DOI：10.16781/j.0258-879x.2017.06.0746 |
| 投稿时间：2017-02-04修订日期：2017-04-10 |
| 基金项目: |
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| Effect of LIM homeobox gene-8 on biological behavior of human ovarian cancer SKOV3 cells |
| HUANG Zheng1,MA Wei1,LU Huan1,WANG Bin1,JIN Guo-hua2,LÜ Guang-ming2,ZHANG Xin-hua2,ZHANG Rui3* |
(1. Department of Obstetrics and Gynecology, Sixth People's Hospital of Shanghai (South Branch), Shanghai Jiaotong University, Shanghai 201499, China;
2. Department of Anatomy, School of Medicine, Nantong University, Nantong 226001, Jiangsu, China;
3. Department of Obstetrics and Gynecology, Sixth People's Hospital of Shanghai, Shanghai Jiaotong University, Shanghai 200233, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the effect of LIM homeobox gene-8 (Lhx8) on the proliferation, metastasis and invasion of human ovarian cancer SKOV3 cells. Methods Lhx8-overexpression lentivirus (LV-Lhx8) was transfected into ovarian cancer SKOV3 cells to establish Lhx8-overexpression model and the negative control lentivirus (LV-NC) was used as control. Then Lhx8-siRNA and FAM-siRNA (used as control; NC group) siRNA was transfected into SKOV3 cells using Lipofectamine 2000 to construct cell interference model. Cells in wild type (WT) group were only given equivalent transfection reagent without virus or any interference fragment. The expression of Lhx8 was detected by immunofluorescence, qPCR and Western blotting. The proliferation of cells after overexpressing or interfering Lhx8 was measured by EDU assays and cell cycle assay. The migration and invasion of cells after transfection were measured by wound scratch experiments and Transwell assay. Results The expression of Lhx8 in SKOV3 cells in the LV-Lhx8 group was significantly higher than that in the LV-NC and WT groups (P<0.01), and its mRNA and protein expressions were significantly decreased after interfering Lhx8 (P<0.01). Compared with the WT and LV-NC groups, the proliferation of SKOV3 cells was significantly decreased in the LV-Lhx8 group and was significantly increased in the Lhx8-siRNA group (P<0.01). The cell cycle assay showed that Lhx8 overexpression significantly inhibited cell proliferation by increasing the number of cells in the G0/G1 phase, while the number of cells in the S phase in the Lhx8-siRNA group was significantly higher than that in the WT and NC groups (P<0.01). The migration and invasion of SKOV3 cells and the expression of matrix metalloproteinase (MMP)-2 and MMP-9 in the LV-Lhx8 group were significantly lower than those in the WT and LV-NC groups (P<0.01), while those in the Lhx8-siRNA group were significantly higher (P<0.01). Conclusion Lhx8 can inhibit the proliferation, migration and invasion of ovarian cancer SKOV3 cells, and down-regulate the expressions of MMP-2 and MMP-9. |
| Key words: ovarian neoplasms LIM homeobox 8 cell proliferation cell migration neoplasm invasiveness |
