结肠癌转移相关基因1对食管癌细胞增殖和迁移的作用及其机制研究

发布时间：2018-03-24 15:38

本文选题：食管鳞状细胞癌　切入点：MACC1　出处：《东南大学》2015年硕士论文

【摘要】：[研究目的]研究结肠癌转移相关基因1 (Metastasis Associated In Colon Cancer 1, MACC1)对食管癌细胞增殖和迁移的作用及可能的机制。[研究方法]1.采用RT-PCR及Western blot检测不同食管癌细胞株EC109、 EC9706、 TE13、 EC1和正常食管上皮细胞株HEEpiC中MACC1的表达,选取低表达MACC1的细胞株TE13和高表达MACC1的细胞株EC109进行后续实验。2.构建携带MACC1的腺病毒载体Ad-MACC1和MACC1 siRNA, Ad-MACC1感染TE13细胞,增加MACC1的表达,MACC1 siRNA转染EC109食管癌细胞,降低MACC1的表达,通过Western blot检测感染或转染效果。3. Ad-MACC1和MACC1 siRNA分别处理TE13和EC109细胞后,通过MTT实验检测MACC1对人食管癌细胞增殖的影响,流式细胞术检测对MACC1细胞周期的影响。采用划痕实验及克隆形成实验观察感染或转染前后食管癌细胞侵袭及迁移变化。4.采用Western blot检测Ad-MACC1和MACC1 siRNA处理食管癌细胞TE13和EC109后的增殖细胞核抗原(PCNA)和c-MET等的表达。[研究结果]1. MACC1在食管癌细胞株EC9706和Eca109中表达较正常食管上皮细胞株HEEpiC中高,而在TE13和EC1中表达较低。2.成功构建Ad-MACC1和MACC1 siRNA, Ad-MACC1感染TE13细胞后,MACC1基因和蛋白的表达增加。而MACC1 siRNA转染EC109细胞后,MACC1基因和蛋白的表达降低。3. Ad-MACC1感染TE13细胞后,细胞的增殖和迁移明显的增加。而MACC1 siRNA转染EC109细胞后,细胞的增殖和迁移受到明显的抑制。4. Ad-MACC1感染TE13细胞后,PCNA和c-MET基因和蛋白表达明显增加,而MACC1 siRNA转染EC109细胞后,PCNA和c-MET基因和蛋白表达较阴性对照组降低。[结论]MACC1在EC9706和Eca109中表达较TE13和EC1高,其高表达可促进细胞的增殖和迁移,而表达下调后明显抑制食管癌细胞的增殖、侵袭及转移潜能,MACC1高表达促进PCNA和c-MET的表达,而低表达抑制其表达。MACC1促进食管癌细胞的增殖、侵袭及转移。其机制可能通过调节下游基因c-MET表达水平实现。
[Abstract]:[objective] to study the effect of Metastasis Associated in Colon Cancer 1 (MACC1) on the proliferation and migration of esophageal carcinoma cells and its possible mechanism. 1. To detect different esophageal cancer cell lines EC109, EC9706, TE13 by RT-PCR and Western blot. Expression of MACC1 in EC1 and normal esophageal epithelial cell line HEEpiC. The cell line TE13 with low expression of MACC1 and the cell line EC109 with high expression of MACC1 were selected for further experiment. 2. The adenovirus vectors Ad-MACC1 and MACC1 siRNAs carrying MACC1 were constructed, Ad-MACC1 infected TE13 cells, and the expression of MACC1 was increased. MACC1 siRNA was transfected into EC109 esophageal carcinoma cells, and the expression of MACC1 was decreased. Western blot was used to detect the effect of infection or transfection. Ad-MACC1 and MACC1 siRNA were used to treat TE13 and EC109 cells respectively. The effect of MACC1 on the proliferation of human esophageal carcinoma cells was detected by MTT assay. The effect of flow cytometry on the cell cycle of MACC1. The invasion and migration of esophageal carcinoma cells before and after infection or transfection were observed by scratch test and clone formation assay. 4. The TE13 of esophageal cancer cells treated with Ad-MACC1 and MACC1 siRNA were detected by Western blot. Expression of proliferating cell nuclear antigen (PCNA) and c-MET after EC109. [results] 1.The expression of MACC1 in EC9706 and Eca109 cells was higher than that in normal esophageal epithelial cell line HEEpiC. Ad-MACC1 and MACC1 siRNAs were successfully constructed in TE13 and EC1. The expression of MACC1 gene and protein increased after Ad-MACC1 infection in TE13 cells, while the expression of MACC1 gene and protein in EC109 cells transfected with MACC1 siRNA decreased .3. Ad-MACC1 infected TE13 cells. The proliferation and migration of EC109 cells were significantly increased after transfection of MACC1 siRNA, and the proliferation and migration of EC109 cells were inhibited significantly. 4. The expression of c-MET gene and protein in TE13 cells infected with Ad-MACC1 was significantly increased. The expression of MACC1 and c-MET gene and protein in EC109 cells transfected with MACC1 siRNA were lower than those in negative control group. [conclusion] the expression of MACC1 in EC9706 and Eca109 was higher than that in TE13 and EC1, and the high expression of MACC1 could promote the proliferation and migration of EC109 cells. The down-regulated expression of MACC1 significantly inhibited the proliferation of esophageal cancer cells, and the high expression of MACC1 promoted the expression of PCNA and c-MET, while the low expression of MACC1 inhibited the proliferation of esophageal cancer cells. Invasion and metastasis. The mechanism may be through regulation of downstream gene c-MET expression level.
【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.1

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