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目的:通过siRNA干扰技术沉默ARTN基因在人食管癌细胞TE11中的表达,研究其对人食管癌细胞增殖、迁移和侵袭能力的影响。方法:设计并合成3条特异性针对ARTN基因的siRNA,构建了真核表达载体pSilencer2.1-ARTN-siRNA,转染到TE11细胞中,经Real-time PCR和Western blot检测ARTN在mRNA和蛋白水平的表达;通过MTT比色法、细胞划痕实验和Boyden小室侵袭实验观察转染pSilencer2.1-ARTN-siRNA后细胞增殖、迁移和侵袭能力的改变。结果:转染重组质粒pSilencer2.1-ARTN-siRNA后人食管癌细胞TE11中ARTN的mRNA表达受到抑制,蛋白表达降低;通过体外实验表明,细胞的增殖能力、迁移能力和侵袭能力均减弱。结论:siRNA沉默ARTN基因的表达可降低人食管癌细胞TE11的增殖、迁移和侵袭能力,ARTN可能是食管癌的一个生物治疗靶点,有望成为治疗食管癌的新途径。
OBJECTIVE: To silence the expression of ARTN gene in human esophageal cancer cell line TE11 by siRNA interference and to study its effect on the proliferation, migration and invasion of human esophageal cancer cells. METHODS: Three siRNAs targeting ARTN gene were designed and synthesized. The eukaryotic expression vector pSilencer2.1-ARTN-siRNA was constructed and transfected into TE11 cells. Real-time PCR and Western blot were used to detect the expression of ARTN mRNA and protein The expression of pSilencer2.1-ARTN-siRNA was detected by MTT assay, cell scratch assay and Boyden chamber invasion assay. The changes of cell proliferation, migration and invasion were observed. Results: The expression of ARTN mRNA in human esophageal cancer cell line TE11 was inhibited and the protein expression was decreased after transfected with recombinant plasmid pSilencer2.1-ARTN-siRNA. In vitro experiments showed that the proliferation, migration and invasion ability of ARTN decreased. Conclusion: The silencing of ARTN gene by siRNA can reduce the proliferation, migration and invasion of human esophageal cancer cell line TE11. ARTN may be a biological therapeutic target for esophageal cancer and may be a new way to treat esophageal cancer.