DLC2在结直肠癌中的表达和对结直肠癌细胞凋亡的影响

齐翀; 洪亮; 李亮; 蔡元坤

doi:10.3969/j.issn.1671-038X.2018.05.08

DLC2在结直肠癌中的表达和对结直肠癌细胞凋亡的影响

- 复旦大学附属上海市第五人民医院, 上海 200240
基金项目:
闵行区自然科学研究课题(No:2015MHZ046)

详细信息

作者简介:
齐翀,男,硕士,副主任医师,研究方向:结肠癌

通讯作者: 齐翀,E-mail:jihao807253@163.com

中图分类号: R735.3

收稿日期: 2017-07-01

Expression of DLC2 in colorectal cancer and its effect on apoptosis of colorectal cancer cells

- Fifth People's Hospital Affiliated to Fudan University, Shanghai 200240, China

More Information

Corresponding author: QI Chong, E-mail: jihao807253@163.com

Received Date: 01 July 2017

摘要

摘要: [目的]探讨肝癌缺失基因2(DCL2)在结直肠癌中的表达及对结直肠癌细胞凋亡的影响。[方法]收集结直肠癌组织及对应的癌旁组织,培养人结直肠癌细胞HCT116、HT29、Caco2和人正常肠上皮细胞FHC,Western blot检测组织及细胞中DLC2表达水平。结直肠癌细胞转染DCL2过表达载体(过表达组)和空载体(阴性组),以只加入转染试剂的细胞为对照组,Western blot检测转染后细胞中DCL2、p38、磷酸化的p38(p-p38)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved cysteinyl aspartate specific proteinase 3,Cleaved Caspase-3)表达,流式细胞术检测细胞凋亡情况。[结果]癌组织中DLC2的表达水平明显低于癌旁组织,差异有统计学意义(t=12.363,P=0.000)。HT29、HCT116、Caco2细胞中DLC2的表达水平明显低于FHC细胞,差异具有统计学意义(t₁=8.624,t₂=9.385,t₂=10.322,P=0.000)。Caco2(0.06±0.03)细胞中DLC2的表达水平明显低于HT29、HCT116,差异具有统计学意义(t₁=8.676,t₂=7.645,P=0.000)。后续实验选用Caco2细胞继续研究。阴性组DLC2的表达水平与对照组相比,差异没有统计学意义(t₁=2.325,P>0.05)。过表达组DLC2的表达水平明显高于对照组,差异具有统计学意义(t₁=13.545,P=0.000)。阴性组细胞凋亡率与对照组相比,差异没有统计学意义(t₁=1.646,P>0.05)。过表达组细胞凋亡率明显高于对照组,差异具有统计学意义(t₁=36.256,P=0.000)。阴性组、过表达组与对照组细胞中p38表达水平没有明显变化(t₁=2.335,t₂=1.464,P>0.05)。阴性组与对照组细胞中p-p38表达水平没有明显变化(t₁=4.587,P>0.05)。阴性组与对照组细胞中Cleaved Caspase-3表达水平没有明显变化(t₁=2.673,P>0.05)。过表达组与对照组细胞中p-p38表达水平相比明显升高,差异有统计学意义(t₁=9.617,P=0.000)。过表达组与对照组细胞中Cleaved Caspase-3表达水平相比明显升高,差异具有统计学意义(t₁=10.696,P=0.000)。[结论]DLC2在结直肠癌中表达下调,DLC2可能通过作用于p38信号通路促进人结直肠癌细胞凋亡。
- 结直肠癌 /
- 凋亡 /
- 肝癌缺失基因2 /
- p38
Abstract: [Objective]To investigate the expression of DCL2 in colorectal carcinoma and its effect on the apoptosis of colorectal cancer cells.[Methods]Colorectal cancer tissues and corresponding adjacent tissues were collected.Then human colorectal cancer cells HCT116,HT29,Caco2 and human normal intestinal epithelial cell line FHC were cultured.After that,the expression of DLC2 in tissues and cells will be detected by Western blot.Colorectal cancer cells were transfected with DCL2 overexpression vector(over expression group)and empty vector(negative group).The cells transfected with transfection reagent were used as control group.Western blot was used to detect the expressions of DCL2,p38,p-p38 and Cleaved Caspase-3 in transfected cells.And flow cytometry will be used to detect cell apoptosis.[Results]The expression level of DLC2 in cancer tissues was significantly lower than that in the adjacent tissues,and the difference was statistically significant(t=12.363,P=0.000).The expression of DLC2 in HT29,HCT116,Caco2 was significantly lower than that of FHC cells,and the difference was statistically significant(t₁=8.624,t₂=9.385,t₂=10.322,P=0.000).The expression level of DLC2 in Caco2 cells was significantly lower than that of HT29 and HCT116,and the difference was statistically significant(t₁=8.676,t₂=7.645,P=0.000).Follow-up experiments were performed by using Caco2 cells.There is no statistically significant difference between the expression level of DLC2 in negative group and that in control group.(t₁=2.325,P＞0.05).The expression level of DLC2 in the over expression group was significantly higher than that in the control group,and the difference was statistically significant(t₁=13.545,P=0.000).There is no statistically significant difference between the apoptosis rate of the negative group and that of control group(t₁=1.646,P＞0.05).The apoptosis rate of over expression group was significantly higher than that of the control group,and the difference was statistically significant(t₁=36.256,P=0.000).Compared with the control group,the expression of p38 in the negative group and over expression group had no significant changes(t₁=2.335,t₂=1.464,P＞0.05).Compared with the control group,the expression of p-p38 in the negative group(0.24±0.04)had no significant change(t₁=4.587,P＞0.05).And there is no significant difference between the expression of Cleaved Caspase-3 in the negative group and the control group(t₁=2.673,P＞0.05).The expression level of p-p38 in the over expression group was significantly higher than that in the control group,and the difference was statistically significant(t₁=9.617,P=0.000).The expression level of Cleaved Caspase-3 in the over expression group was significantly higher than that in the control group,and the difference was statistically significant(t₁=2.673,P＞0.05).[Conclusion]There is downregulation of DLC2 expression in colorectal cancer cells.DLC2 may promote apoptosis of human colorectal cancer cells by acting on p38 signaling pathway.
- colorectal cancer /
- apoptosis /
- deleted in liver cancer 2 /
- p38

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