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[目的]研究nectin 4在肺癌组织中的表达及其在肺癌细胞增殖、迁移侵袭过程中的作用。[方法]免疫组化法检测20例肺癌组织及其对应癌旁组织中nectin 4的表达情况;QPCR及Western Blot分别检测不同肺癌细胞株及正常肺上皮细胞中nectin 4 mRNA和蛋白的表达情况;设计nectin 4干扰RNA序列,转染肺癌细胞后,MTT法检测细胞的增殖能力。划痕愈合实验和Transwell小室法分别检测细胞的迁移能力和侵袭能力。[结果]免疫组化结果显示nectin 4在癌旁组织中不表达,而40%的肺癌组织中表达阳性(8/20),且nectin 4的表达与肺癌患者淋巴结转移明显相关(r=0.612,P=0.004)。QPCR及Western Blot结果显示,正常肺上皮细胞BEAS-2B中nectin 4表达较低,而在A549和PC-9肺癌细胞株中nectin 4表达明显较高。设计的其中一条nectin 4 siRNA可以降低肺癌A549和PC-9细胞nectin 4蛋白表达70%以上,且nectin 4基因干扰后,肺癌细胞的增殖能力减弱,迁移和侵袭能力也受到明显抑制。[结论]Nectin 4表达水平与肺癌细胞的增殖、迁移和侵袭密切相关,可以作为肺癌转移治疗的重要靶点。
[Objective] To investigate the expression of nectin 4 in lung cancer and its role in the proliferation, migration and invasion of lung cancer cells. [Methods] The expression of nectin 4 in 20 cases of lung cancer tissues and corresponding paracancerous tissues was detected by immunohistochemistry. The expression of nectin 4 mRNA and protein in different lung cancer cell lines and normal lung epithelial cells were detected by QPCR and Western Blot. Design nectin 4 interference RNA sequence, transfected lung cancer cells, MTT assay of cell proliferation. Scratch healing assay and Transwell chamber method were used to detect cell migration and invasion ability. [Results] The results of immunohistochemistry showed that nectin 4 was not expressed in adjacent tissues and 40% of lung cancer tissues (8/20), and the expression of nectin 4 was significantly correlated with lymph node metastasis in lung cancer patients (r = 0.612, P = 0.004). The results of QPCR and Western Blot showed that the expression of nectin 4 in normal lung epithelial cells was lower than that in normal lung epithelial cells and nectin 4 was significantly higher in A549 and PC-9 lung cancer cell lines. One of the designed nectin 4 siRNAs can reduce the expression of nectin 4 protein in lung cancer A549 and PC-9 cells by more than 70%. The proliferation of lung cancer cells is attenuated by nectin 4 gene knockdown, and migration and invasion ability are also significantly inhibited. [Conclusion] The expression level of Nectin 4 is closely related to the proliferation, migration and invasion of lung cancer cells and may be used as an important target for lung cancer metastasis.