miR-200b通过促进NKD2调节子宫内膜癌细胞EMT及对子宫内膜癌侵袭和迁移能力的影响

miR-200b regulates the EMT of endometrial cancer cells by promoting NKD2 and affects the invasion and migration of endometrial cancer

目的 探讨miR-200b通过促进NKD2调节子宫内膜癌细胞EMT及对子宫内膜癌侵袭和迁移能力的影响。方法 采用qRT-PCR检测组织中miR-200b和NKD2的表达,免疫组化染色检测组织中NKD2的阳性表达,Transwell实验检测细胞侵袭,细胞划痕实验检测细胞迁移,Westernblot检测EMT相关蛋白及NKD2表达,NKD2过表达载体构建,检测EMT相关蛋白及NKD2表达。结果 子宫内膜腺癌组织中miR-200b和NKD2的表达均低于癌旁组织(P<0.001);子宫内膜腺癌组织中NKD2的阳性率明显高于癌旁组织(P<0.001);miR-200b模拟物组中HEC-1A细胞的侵袭数量和迁移能力较阴性对照组相比明显降低(P<0.001),miR-200b抑制组中HEC-1A细胞的侵袭数量和迁移能力较阴性对照组相比显著升高(P<0.001);miR-200b模拟物组中E-cadherin和NKD2蛋白水平明显高于阴性对照组(P<0.001),miR-200b模拟物组中N-cadherin和Snail蛋白水平明显低于阴性对照组(P<0.001);miR-200b抑制组中E-cadherin和NKD2蛋白水平显著低于阴性对照组(P<0.001),miR-200b模拟物组中N-cadherin和Snail蛋白水平显著高于阴性对照组(P<0.001);miR-200b mimic+pcDNA-NKD2组中E-cadherin和NKD2蛋白水平明显高于miR-200b mimic组(P<0.001),miR-200b mimic+pcDNA-NKD2组中N-cadherin和Snail蛋白水平明显低于miR-200b mimic组(P<0.001)。结论 miR-200b通过促进NKD2的表达抑制子宫内膜癌细胞EMT,并可能通过EMT调节宫内膜癌侵袭和迁移能力。

Objective To investigate the effect of miR-200b on the invasiveness and migration of endometrial cancer cells by promoting NKD2 to regulate EMT of endometrial cancer cells. Methods qRT-PCR was used to detect the expression of miR-200b and NKD2 in the tissue, immunohistochemical staining was used to detect the positive expression of NKD2 in the tissue, Transwell assay was used to detect cell invasion, cell scratch assay was used to detect cell migration, and Western blot was used to detect the expression of EMT-related proteins and NKD2. NKD2 overexpression vector was constructed, and EMT-related proteins and NKD2 expression were detected. Results The expressions of miR-200b and NKD2 in endometrial adenocarcinoma tissues were lower than those in paracancerous tissues(P<0.001). The positive rate of NKD2 in endometrial adenocarcinoma tissues was significantly higher than that in paracancerous tissues(P<0.001). The invasion number and migration ability of HEC-1A cells in the miR-200b mimic group were significantly lower than those in the negative control group(P<0.001). The protein levels of E-cadherin and NKD2 in the miR-200b mimic group were significantly higher than those in the negative control group(P<0.001), and the protein levels of N-cadherin and Snail in the miR-200b mimic group The levels of E-cadherin and NKD2 in the miR-200b inhibition group were significantly lower than those in the negative control group(P<0.001). The protein levels of N-cadherin and Snail in the miR-200b mimic group were significantly higher than those in the negative control group(P<0.001). The protein levels of E-cadherin and NKD2 in the miR-200b mimic+pcDNA-NKD2 group were significantly higher than those in the miR-200b mimic group. The protein levels of N-cadherin and Snail in the miR-200b mimic+pcDNA-NKD2 group were significantly lower than those in the miR-200b mimic group(P<0.001). Conclusion miR-200b inhibits EMT of endometrial cancer cells by promoting the expression of NKD2, and may regulate the invasion and migration ability of endometrial cancer through EMT.