溶酶体组织蛋白酶L对人卵巢癌SKOV3细胞迁移及侵袭的作用

发布时间：2018-06-13 08:28

本文选题：溶酶体组织蛋白酶L + 上皮-间充质转化　；参考：《肿瘤防治研究》2015年06期

【摘要】：目的探讨溶酶体组织蛋白酶L(Cathepsin L)是否通过上皮-间充质转化(EMT)影响卵巢癌细胞的侵袭及迁移能力。方法荧光定量PCR法检测人卵巢癌细胞ES-2、SKOV3、OV1以及OV2中Cathepsin L的表达水平;设计并合成靶向Cathepsin L的特异性sh RNA,通过脂质体转染法转染Cathepsin L表达最高的卵巢癌细胞SKOV3以构建稳定低表达Cathepsin L细胞株,Western blot和定量PCR法验证sh RNA的干扰效率;划痕实验及Transwell法检测干扰后细胞的迁移及侵袭能力,Western blot法检测EMT相关指标E-cadherin和N-cadherin以及其上游信号分子Snail、p-AKT的变化。结果四株卵巢癌细胞中,SKOV3的Cathepsin L表达水平最高(以此为参照),而ES-2、OV1以及OV2细胞的表达水平分别为0.72±0.04、0.34±0.03和0.55±0.05。Cathepsin L-sh RNA转染SKOV3细胞后,SKOV3/sh RNA中的Cathepsin L的表达水平较空白组和对照组显著下降;划痕12 h和24 h后,SKOV3/sh RNA组的细胞迁移能力明显受到抑制;SKOV3、SKOV3/Con和SKOV3/sh RNA组的穿膜细胞数分别为(93.67±8.62)、(90.33±12.22)、(35.67±4.73),与对照组相比,SKOV3/sh RNA组细胞侵袭能力受到显著抑制(P0.01);Cathepsin L干扰组细胞的E-cadherin表达增加,N-cadherin的表达降低;此外,Cathepsin L干扰组细胞的Snail、p-AKT表达较对照组显著下降。结论 Cathepsin L可以促进卵巢癌细胞的迁移和侵袭;其机制可能与调节EMT的上游信号分子Snail、p-AKT有关。提示Cathepsin L在卵巢癌的发生发展中起重要作用。
[Abstract]:Objective to investigate whether lysosomal cathepsin L (lysosomal cathepsin L) affects the invasion and migration of ovarian cancer cells through epithelial-mesenchymal transformation (EMTT). Methods fluorescence quantitative PCR was used to detect the expression of Cathepsin L in human ovarian cancer cell line ES-2SKOV3OV1 and OV2. The specific sh RNAs targeting Cathepsin L were designed and synthesized. The ovarian cancer cell line SKOV3, which had the highest expression of Cathepsin L, was transfected by liposome transfection to construct a cell line with stable and low expression of Cathepsin L. Western blot and quantitative PCR were used to verify the interference efficiency of sh RNA. The migration and invasiveness of EMT-related markers E-cadherin, N-cadherin and its upstream signal molecule Snail-p-AKT were detected by scratch test and Transwell method. The changes of E-cadherin and N-cadherin were detected by Western blot. Results the expression level of Cathepsin L in SKOV3 cells was the highest (as compared with the control group). The expression levels of Cathepsin L in SKOV1 and OV2 cells were 0.72 卤0.04, 0.34 卤0.03 and 0.55 卤0.05.Cathepsin L-sh RNA transfection into SKOV3 cells, respectively. After 12 and 24 hours of scratch, the cell migration ability of SKOV3 / RSH RNA group was significantly inhibited. The number of perforated cells in SKOV3 / SKOV3 / Con and SKOV3 / sh RNA group was 93.67 卤8.62 卤12.22 ~ 0.33 卤12.22 ~ 0.33 卤12.22 ~ 3.73, respectively. Compared with the control group, the cell invasion ability of SKOV3 / sh RNA group was significantly inhibited by P0.01Cathepsin L interfering E-cadherin table. The expression of N-cadherin decreased. In addition, the expression of Snail-p-AKT in Cathepsin L interference group was significantly lower than that in control group. Conclusion Cathepsin L can promote the migration and invasion of ovarian cancer cells, and its mechanism may be related to the up-stream signal molecule Snail-p-AKT which regulates EMT. These results suggest that Cathepsin L plays an important role in the development of ovarian cancer.
【作者单位】：广州医科大学附属第三医院生殖中心;南方医科大学肿瘤中心;
【基金】：广州市科技计划项目(民主科技重大专项)(2012Y2-00022) 广东省自然科学基金博士启动项目(S2013040013849) 广州医科大学校基金博士启动项目(2012C54) 南方医科大学中西医结合医院博士启动基金(1201302009)
【分类号】：R737.31

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