阻断Wnt信号通路对人绒毛膜癌细胞株JEG-3迁移及侵袭能力的影响
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摘要
目的观察阻断Wnt信号通路后,人绒毛膜癌细胞株JEG-3细胞迁移及侵袭能力的影响。方法将人绒毛膜癌细胞株JEG-3细胞传代后,按完全随机法分为实验组和对照组,每组6孔。实验组在常规细胞培养基础上,给予外源性Wnt信号通路阻断剂DKK-1(100 ng/mL),对照组加入同体积细胞培养基。运用蛋白印迹法(Western blot法)、逆转录聚合酶链反应(RTPCR)法检测两组细胞中β-catenin、Wnt、E-cadherin、Vimentin等蛋白及mRNA的表达;采用划痕实验、Transwell实验观测两组细胞迁移及侵袭能力的变化。结果与对照组相比,实验组β-catenin、Wnt、Vimentin蛋白及mRNA的表达均降低,E-cadherin蛋白及mRNA的表达增高,差异均有统计学意义(P<0.05);实验组肿瘤细胞迁移及穿膜细胞数较对照组均降低,差异有统计学意义(P<0.05)。结论阻断Wnt信号通路可抑制肿瘤细胞上皮-间充质转化过程,进而降低人绒毛膜癌细胞株JEG-3细胞的迁移及侵袭能力。
Objective To observe the changes of migration and invasion capacity in the human choriocarcinoma cell line JEG-3 after blockade of Wnt signal pathway. Methods The subculturing cells of human choriocarcinoma cell line JEG-3 were divided into the experiment group and the control groupby means of complete randomization,6 holes in each group. Besides routine cell culture for the two groups,the experiment group was treated with additional exogenous Wnt signal pathway blocker DKK-1( 100 ng/mL),and the control group with cell culture media of same volume. The protein and mRNA expression of beta-catenin,Wnt,E-cadherin andvimentin in the cells of both groupswere evaluated by means of reverse transcriptase-polymerase chain reaction( RT-PCR) and western blotting,and changes of migration and invasion capacity in these choriocarcinoma cells were examined by means of scratch test and transwell assay. Results Compared with the control group,the protein and mRNA expression of beta-catenin,Wnt and vimentin in the experiment group decreased evidently,but the protein and mRNA expression of E-cadherin increased,and differences between the two groups were all of statistical significance( P < 0. 05). The number of tumor cells migrated and transported across membrane in the experiment group was significantly lower than that in the control group( P < 0. 05). Conclusion Blocking the Wnt signal pathway would inhibit the process of epithelial-mesenchymal transition( EMT) inchoriocarcinoma cells,and further decrease the migration and invasion capacity of human choriocarcinoma cell line JEG-3.
Objective To observe the changes of migration and invasion capacity in the human choriocarcinoma cell line JEG-3 after blockade of Wnt signal pathway. Methods The subculturing cells of human choriocarcinoma cell line JEG-3 were divided into the experiment group and the control groupby means of complete randomization,6 holes in each group. Besides routine cell culture for the two groups,the experiment group was treated with additional exogenous Wnt signal pathway blocker DKK-1( 100 ng/mL),and the control group with cell culture media of same volume. The protein and mRNA expression of beta-catenin,Wnt,E-cadherin andvimentin in the cells of both groupswere evaluated by means of reverse transcriptase-polymerase chain reaction( RT-PCR) and western blotting,and changes of migration and invasion capacity in these choriocarcinoma cells were examined by means of scratch test and transwell assay. Results Compared with the control group,the protein and mRNA expression of beta-catenin,Wnt and vimentin in the experiment group decreased evidently,but the protein and mRNA expression of E-cadherin increased,and differences between the two groups were all of statistical significance( P < 0. 05). The number of tumor cells migrated and transported across membrane in the experiment group was significantly lower than that in the control group( P < 0. 05). Conclusion Blocking the Wnt signal pathway would inhibit the process of epithelial-mesenchymal transition( EMT) inchoriocarcinoma cells,and further decrease the migration and invasion capacity of human choriocarcinoma cell line JEG-3.
引文
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[11]提爱军,向丹,李磊华,等.9-顺式视黄酸抑制人胆管癌细胞株QBC939Wnt信号通路和细胞迁移[J].中华实验外科杂志,2015,32(11):2632-2635.
[12]陈海滔,徐超,姚庆华.姜黄素通过调节Wnt/β-catenin信号通路抑制结肠癌细胞上皮间质转化[J].肿瘤学杂志,2017,23(1):35-39.
[13]邹慧娟,郑航,李明君,等.Wnt信号通路在结肠癌中的表达及其作用机制[J].中国老年学杂志,2016,36(14):3373-3374.
[2]PRASAD C P,RATH G,MATHUR S,et al.Expression analysis of E-cadherin,Slug and GSK3 beta in invasive ductal carcinoma of breast[J].BMC Cancer,2009,9(1):325.
[3]KAUFHOLD S,BONAVIDA B.Central role of Snail1 in the regulation of EMT and resistance in cancer:a target for therapeutic intervention[J].J Exp Clin Cancer Res,2014,33(1):62.
[4]HUANG D,DU X.Crosstalk between tumor cells and microenvironment via Wnt pathway in colorectal cancer dissemination[J].World J Gastroenterol,2008,14(12):1823-1827.
[5]包秀丽,刘婷婷,王艳艳,等.Wnt3a诱导人晶状体上皮细胞细胞外基质合成和胶原凝胶收缩的机制研究[J].中华实验眼科杂志,2016,34(7):597-601.
[6]张延新,赵红军,宋文刚,等.TWEAK反义寡核苷酸对人结肠癌细胞系SW480增殖及侵袭能力的影响[J].解剖学报,2014,45(1):74-79.
[7]郝淑玲,王卫平,李建东,等.肾上腺髓质素和转化生长因子β1对肺成纤维细胞增殖及c-myc表达的影响[J].中华实用诊断与治疗杂志,2015,29(4):333-335.
[8]王科峰,柴林燕.替米沙坦对AGEs诱导人内皮细胞表达VCAM-1及MCP-1的作用和机制[J].安徽医学,2017,38(4):395-400.
[9]王丽,张宁,张龙江,等.SCUBE2过表达通过Wnt/β-catenin抑制结直肠癌细胞上皮-间质转化[J].中国病理生理学杂志,2016,32(12):2245-2250.
[10]李琳,徐莉,范秋灵,等.熊果酸通过调节Wnt/β-catenin通路抑制高糖诱导的足细胞上皮间充质转分化[J].中华肾脏病杂志,2016,32(5):365-370.
[11]提爱军,向丹,李磊华,等.9-顺式视黄酸抑制人胆管癌细胞株QBC939Wnt信号通路和细胞迁移[J].中华实验外科杂志,2015,32(11):2632-2635.
[12]陈海滔,徐超,姚庆华.姜黄素通过调节Wnt/β-catenin信号通路抑制结肠癌细胞上皮间质转化[J].肿瘤学杂志,2017,23(1):35-39.
[13]邹慧娟,郑航,李明君,等.Wnt信号通路在结肠癌中的表达及其作用机制[J].中国老年学杂志,2016,36(14):3373-3374.