本文選題：溶酶體組織蛋白酶L + 上皮-間充質(zhì)轉(zhuǎn)化�。� 參考：《腫瘤防治研究》2015年06期

【摘要】：目的探討溶酶體組織蛋白酶L(Cathepsin L)是否通過(guò)上皮-間充質(zhì)轉(zhuǎn)化(EMT)影響卵巢癌細(xì)胞的侵襲及遷移能力。方法熒光定量PCR法檢測(cè)人卵巢癌細(xì)胞ES-2、SKOV3、OV1以及OV2中Cathepsin L的表達(dá)水平;設(shè)計(jì)并合成靶向Cathepsin L的特異性sh RNA,通過(guò)脂質(zhì)體轉(zhuǎn)染法轉(zhuǎn)染Cathepsin L表達(dá)最高的卵巢癌細(xì)胞SKOV3以構(gòu)建穩(wěn)定低表達(dá)Cathepsin L細(xì)胞株,Western blot和定量PCR法驗(yàn)證sh RNA的干擾效率;劃痕實(shí)驗(yàn)及Transwell法檢測(cè)干擾后細(xì)胞的遷移及侵襲能力,Western blot法檢測(cè)EMT相關(guān)指標(biāo)E-cadherin和N-cadherin以及其上游信號(hào)分子Snail、p-AKT的變化。結(jié)果四株卵巢癌細(xì)胞中,SKOV3的Cathepsin L表達(dá)水平最高(以此為參照),而ES-2、OV1以及OV2細(xì)胞的表達(dá)水平分別為0.72±0.04、0.34±0.03和0.55±0.05。Cathepsin L-sh RNA轉(zhuǎn)染SKOV3細(xì)胞后,SKOV3/sh RNA中的Cathepsin L的表達(dá)水平較空白組和對(duì)照組顯著下降;劃痕12 h和24 h后,SKOV3/sh RNA組的細(xì)胞遷移能力明顯受到抑制;SKOV3、SKOV3/Con和SKOV3/sh RNA組的穿膜細(xì)胞數(shù)分別為(93.67±8.62)、(90.33±12.22)、(35.67±4.73),與對(duì)照組相比,SKOV3/sh RNA組細(xì)胞侵襲能力受到顯著抑制(P0.01);Cathepsin L干擾組細(xì)胞的E-cadherin表達(dá)增加,N-cadherin的表達(dá)降低;此外,Cathepsin L干擾組細(xì)胞的Snail、p-AKT表達(dá)較對(duì)照組顯著下降。結(jié)論 Cathepsin L可以促進(jìn)卵巢癌細(xì)胞的遷移和侵襲;其機(jī)制可能與調(diào)節(jié)EMT的上游信號(hào)分子Snail、p-AKT有關(guān)。提示Cathepsin L在卵巢癌的發(fā)生發(fā)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 鹵0.04, 0.34 鹵0.03 and 0.55 鹵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 鹵8.62 鹵12.22 ~ 0.33 鹵12.22 ~ 0.33 鹵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.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第三醫(yī)院生殖中心;南方醫(yī)科大學(xué)腫瘤中心;
【基金】：廣州市科技計(jì)劃項(xiàng)目(民主科技重大專項(xiàng))(2012Y2-00022) 廣東省自然科學(xué)基金博士啟動(dòng)項(xiàng)目(S2013040013849) 廣州醫(yī)科大學(xué)�；鸩┦繂�(dòng)項(xiàng)目(2012C54) 南方醫(yī)科大學(xué)中西醫(yī)結(jié)合醫(yī)院博士啟動(dòng)基金(1201302009)
【分類(lèi)號(hào)】：R737.31

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