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目的:探讨肺腺癌转移相关转录因子1(metastasis-associated lungadenocarcinoma transcript 1,MALAT1)对口腔鳞状细胞癌侵袭能力的影响。方法:应用RT-PCR方法检测MALAT1在口腔鳞状细胞癌组织标本、正常口腔黏膜组织标本以及口腔鳞癌细胞系中的表达;利用小干扰RNA(si RNA)敲低MALAT1在人舌鳞状细胞癌细胞Tscca中的表达;MTT法检测细胞的增殖能力变化;划痕实验、Transwell实验检测肿瘤细胞迁移、侵袭能力的变化;蛋白质印迹法检测肿瘤细胞迁移、侵袭及上皮间质转化(epithelial-mesenchymal transition,EMT)等相关蛋白的表达变化;免疫荧光法检测细胞EMT相关蛋白表达变化;建立Tscca裸鼠皮下荷瘤模型,免疫组织化学染色法检测细胞增殖、侵袭相关蛋白表达。结果:MALAT1在口腔鳞癌组织中的表达明显高于正常组织。抑制MALAT1表达后细胞增殖率下降,细胞系划痕闭合减慢,通过Transwell小室聚碳酸酯膜的细胞数减少,与对照组比较,差异有统计学意义(P<0.05);基质金属蛋白酶-2、-9(MMP-2、MMP-9)、神经钙黏素(N-cadherin)蛋白表达水平明显下调,钙黏着蛋白(E-cadherin)表达水平上调。免疫荧光显示,细胞神经钙黏素荧光强度明显减弱,钙黏着蛋白荧光强度显著增强。体内实验结果显示,MALAT1 si RNA治疗组裸鼠皮下荷瘤体积小于空白对照组及无义序列组(F=18.664,P<0.001);免疫组织化学染色结果示,MALAT1 si RNA治疗组中增殖核抗原(PCNA)、MMP-2、MMP-9表达减少。结论:MALAT1在口腔鳞癌组织中过表达,敲低人舌鳞癌细胞中MALAT1的表达可抑制舌鳞癌细胞的迁移、侵袭能力,MALAT1可能通过调控EMT促进口腔鳞癌的增殖和侵袭过程。
Objective: To investigate the effect of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on the invasiveness of oral squamous cell carcinoma. Methods: The expression of MALAT1 in oral squamous cell carcinoma tissues, normal oral mucosa tissues and oral squamous cell carcinoma cell lines was detected by RT-PCR. The expression of MALAT1 in human tongue squamous cells The changes of cell proliferation were detected by MTT assay. The scratch test, the changes of tumor cell migration and invasion were detected by Transwell assay. The migration, invasion and epithelial-mesenchymal transition of tumor cells were detected by Western blotting. transition and EMT). The expression of EMT-related proteins was detected by immunofluorescence. The subcutaneous tumor-bearing model of Tscca nude mice was established. The proliferation and invasion-related proteins were detected by immunohistochemical staining. Results: The expression of MALAT1 in oral squamous cell carcinoma was significantly higher than that in normal tissues. Compared with the control group, the cell proliferation rate of MALAT1 was decreased, the scratch of cell lines was slowed down, and the number of cells passing through Transwell chamber polycarbonate membrane was decreased. The difference was statistically significant (P <0.05); the expression of matrix metalloproteinase-2 , -9 (MMP-2, MMP-9), N-cadherin protein expression and E-cadherin expression were up-regulated. Immunofluorescence showed that the fluorescence intensity of cadherin decreased significantly and the fluorescence intensity of cadherin significantly increased. The results of in vivo experiments showed that the tumor volume of MALAT1 si RNA treatment group was smaller than that of blank control group and nonsense sequence group (F = 18.664, P <0.001). The results of immunohistochemical staining showed that MALAT1 si RNA treatment group Antigen (PCNA), MMP-2, MMP-9 expression decreased. Conclusion: MALAT1 is overexpressed in oral squamous cell carcinoma. MALAT1 knockdown in human tongue squamous cell carcinoma can inhibit the migration and invasion of tongue squamous cell carcinoma. MALAT1 may promote the proliferation and invasion of oral squamous cell carcinoma by regulating EMT.