LYPD1通过Akt通路促进卵巢癌恶性行为的实验研究

Experimental study of LYPD1 promoting malignant behavior of ovarian cancer through Akt pathway

目的探讨卵巢癌组织中LYPD1表达以及其对卵巢癌恶性生物学行为的影响和可能作用机制。方法采用GEPIA和Oncomine数据库分析正常对照组织和卵巢癌组织中LYPD1 mRNA的表达水平,采用GEPIA网站在线分析LYPD1表达与卵巢癌患者预后的关系。采用实时荧光定量PCR(q PCR)检测卵巢癌组织和癌旁正常组织中LYPD1 mRNA表达。采用CCK-8方法检测LYPD1表达对卵巢癌A2780细胞增殖的影响,采用流式细胞术检测LYPD1表达对A2780细胞周期分布的影响。Western blotting检测LYPD1对ERK和Akt信号通路的影响。结果 Oncomine、TCGA数据库均显示卵巢癌组织中LYPD1的表达水平明显高于正常对照组,差异具有统计学意义(P<0. 01)。q PCR结果显示,在癌旁正常组织、Ⅰ+Ⅱ期、Ⅲ+Ⅳ期卵巢癌组织中LYPD1 mRNA的表达水平分别为1. 07±0. 63、5. 24±1. 59、8. 14±1. 94,3组间差异具有统计学意义(P<0. 05)。GEPIA网站在线分析结果显示,LYPD1表达与卵巢癌预后无关,差异无统计学意义(HR=0. 85,P=0. 2)。PCMVLYPD1组24、48、72、96 h的细胞增殖活性均高于对照组;而LYPD1 shRNA组24、48、72、96 h的细胞增殖活性均低于对照组。PCMV-LYPD1组G1期细胞比率高于对照组[(72. 08±3. 16)%vs.(58. 53±2. 82)%],差异具有统计学意义(P<0. 05)。沉默LYPD1表达提高Akt磷酸化水平(P<0. 05)。结论 LYPD1通过Akt信号途径调控卵巢癌恶性进展。

Objective To investigate the expression of LYPD1 in ovarian cancer and its effect on the biological behavior of ovarian cancer.Methods The expression levels of LYPD1 mRNA in normal tissues and ovarian cancer tissues were analyzed using GEPIA and Oncomine databases,and the relationship between LYPD1 expression and the prognosis of ovarian cancer patients was analyzed using GEPIA website.The expression of LYPD1 mRNA in ovarian cancer and paracancerous normal tissues was detected by real-time fluorescent quantitative PCR(qPCR).The effect of LYPD1 expression on the proliferation of ovarian cancer A2780 cells was detected by CCK-8 method,and the effect of LYPD1 expression on the cell cycle distribution of A2780 cells was detected by flow cytometry.Western blotting was used to detect the effects of LYPD1 on ERK and Akt signaling pathways.Results Both the Oncomine and TCGA databases showed that LYPD1 expression in ovarian cancer tissues was significantly higher than that in normal control group,with statistically significant differences(P<0.01).The results of qPCR showed that the expression of LYPD1 mRNA was 1.07±0.63,5.24±1.59 and 8.14±1.94 in paracancerous normal tissues,stage I+II and stage III+IV ovarian cancer tissues,respectively.The difference between the three groups was statistically significant(P<0.05).The online analysis by GEPIA showed that LYPD1 expression was not associated with the prognosis of ovarian cancer,and the difference was not statistically significant(H r=0.85,P=0.2).The cell proliferation activity of PCMV-LYPD1 group at 24,48,72 and 96 h was higher than that of the control group.The cell proliferation activity of LYPD1 shRNA group at 24,48,72 and 96 h was lower than that of control group.The G 1 phase cell ratio of PCMV-LYPD1 group was higher than that of the control group[(72.08±3.16)%vs.(58.53±2.82)%],and the difference was statistically significant(P<0.05).Silencing LYPD1 expression increased Akt phosphorylation(P<0.05).Conclusion LYPD1 regulates malignant progression of ovarian cancer through the Akt signaling pathway.