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目的:研究S期激酶相关蛋白2(Skp2)对大肠癌侵袭和转移的影响。方法:采用免疫组化SP法检测35例大肠癌及15例癌旁组织中Skp2、基质金属蛋白酶2(MMP2)及基质金属蛋白酶9(MMP9)的表达水平,并分析它们与大肠癌临床病理特征之间的关系;构建Skp2干扰腺病毒与转录因子Sp1过表达腺病毒,转染人结肠癌SW480细胞;用Western blot检测蛋白表达情况;划痕实验检测细胞迁移能力;侵袭实验检测细胞侵袭能力变化。结果:免疫组化结果显示大肠癌中Skp2、MMP2和MMP9阳性表达率分别为(35.5±12.9)%、(78.8±11.8)%、(75.3±11.6)%(P<0.05);Skp2阳性表达率与淋巴结转移及肿瘤分化程度正相关(P<0.05);MMP2、MMP9阳性表达率与肿瘤TNM分期、淋巴结转移正相关(P<0.05),大肠癌中Skp2与MMP2、MMP9表达分别呈正相关(r=0.341,P=0.045;r=0.339,P=0.046)。沉默Skp2后,MMP2、MMP9、Sp1蛋白明显下调(P<0.05);伤口愈合能力明显降低,侵袭细胞数明显减少(P<0.05);加入Sp1过表达腺病毒上调Sp1蛋白后,细胞侵袭能力恢复。结论:Skp2、MMP-2和MMP-9与大肠癌的进展和转移密切相关;Skp2-shRNA可能通过下调Sp1减少MMP2、MMP9的表达从而抑制大肠癌细胞SW480的侵袭和转移。
Objective: To investigate the effect of S-phase kinase-related protein 2 (Skp2) on the invasion and metastasis of colorectal cancer. Methods: The expressions of Skp2, MMP2 and MMP9 in 35 cases of colorectal carcinoma and 15 cases of paracancerous tissues were detected by immunohistochemical SP method and their correlation with clinicopathological features The expression of Skp2 was detected by Western blot. The cell migration ability was detected by scratch assay. The invasiveness of cells was detected by flow cytometry . Results: The positive rates of Skp2, MMP2 and MMP9 in colorectal carcinoma were (35.5 ± 12.9)%, (78.8 ± 11.8)% and (75.3 ± 11.6)%, respectively (P <0.05). The positive expression rate of MMP2 and MMP9 was positively correlated with TNM stage and lymph node metastasis (P <0.05), while there was a positive correlation between Skp2 and MMP2 and MMP9 in colorectal carcinoma (r = 0.341, P = 0.045; r = 0.339, P = 0.046). After silencing Skp2, MMP2, MMP9 and Sp1 protein were significantly downregulated (P <0.05), wound healing was significantly reduced, and the number of invasive cells was significantly decreased (P <0.05). Sp1 protein was up-regulated by Sp1 overexpression adenovirus and cell invasion was restored . Conclusion: Skp2, MMP-2 and MMP-9 are closely related to the progression and metastasis of colorectal cancer. Skp2-shRNA may inhibit the invasion and metastasis of colorectal cancer SW480 by down-regulating Sp1 and decreasing the expression of MMP2 and MMP9.