WWC3下调Snail和N-cadherin,上调E-cadherin抑制非小细胞肺癌侵袭和转移

WWC3 inhibits the invasion and metastasis of non-small cell lung cancer by Snail and N-cadherin down regulation and E-cadherin up regulation

目的研究WWCs(WW and C2 Domain containing protein family)及其各亚型(WWC1,WWC2,WWC3)在人类肺癌中的表达是否存在差异、与临床病理因素的关系及其发挥生物学作用的相关机制。方法应用免疫组化方法检测了159例非小细胞肺癌组织标本中WWCs蛋白表达和亚细胞定位,并对比分析与上皮间质转化相关蛋白E-cadherin、N-cadherin表达的关系。利用集落形成实验和transwell实验,检测WWC3对肺癌细胞增殖及侵袭能力的影响。转染或干扰WWC3的表达后,通过Western blot检测双向调控WWC3对E-cadherin、N-cadherin及Snail表达水平的影响。结果 WWC3在正常支气管上皮细胞浆中高表达(31/40,77.5%),而在非小细胞肺癌中低表达(70/159,44%),明显低于正常支气管上皮的阳性表达率(P<0.001),且其低表达与非小细胞肺癌的低分化(P=0.010)、高TNM分期(P=0.016)、淋巴结转移(P=0.002)相关。WWC3的低表达与上皮间质转化相关蛋白E-cadherin的高表达(P=0.032)、N-cadherin的低表达(P=0.007)相关。在H1299细胞中转染WWC3表达质粒,抑制肺癌细胞的增殖与侵袭能力,下调Snail和N-cadherin,上调E-cadherin的表达。在A549细胞中转染WWC3-si RNA能够明显促进肺癌细胞的增殖和侵袭能力,上调Snail和N-cadherin的表达,而下调E-cadherin的表达。结论 WWC3抑制肺癌细胞侵袭和转移可能与下调Snail和N-cadherin,上调E-cadherin的表达相关。

Objective To study the expressions and clinicopathological significance of WWCs （WW and C2 domain containing） and the subtypes（WWC1,WWC2,WWC3） in bmnan non-small-cell lung cancer. Methods The Immunohistochemical staining was used to check WWCs protein expression in human non-small-cell lung cancer. Colony forming experiment and transwell experiment were used to check WWCs influence on lung cancer cell proliferation and invasion. Western blot was used to check the expression of E-cadherin, N-cadherin and Snail. Results WWC3 was expressed highly in normal bronchial cytoplasm（31/40, 77.5%）, lower in lung cancer cells （70/159, 44%, P〈0.001）, and related to low differentiation （P=0.010） and high TNM stage （P=0.016） and lymph node metastasis（P=0.002）, also related to the expression of E-cadherin（P=0.032） and N-cadherin（P=0.007）. WWC3 expression plasmid transfection inhibited the proliferation and invasion ability of H1299 cells, downregnlated Snail and N-cadherin expression, upregulated E-cadherin expression; but transfected WWC3-siRNA promoted the proliferation and invasion of A549 cells, upregulated Snail and N- cadherin expression, downregulated E-cadherin expression. Conclusion The inhibition of WWC3 on the invasion and metastasis of lung cancer cells might be related to down-regulating the expression of Snail and N-cadherin, up-regulating the expression of E-cadherin.