水通道蛋白5在体外调节结肠癌细胞生长中的作用
[Role of aquaporin-5 in regulating colorectal cancer cell growth in vitro].
作者信息
Chen Shuo, Shan Tao, Wang Li, Chen Xi, Cui Xi-Juan, Gao Xian-Ni
机构信息
Department of General Surgery, Second Affiliated Hospital, Xi'an Jiaotong University, Xi'an710004 China. E-mail:
出版信息
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Oct 20;37(10):1330-1336. doi: 10.3969/j.issn.1673-4254.2017.10.08.
OBJECTIVE
To study the role of aquaporin 5(AQP5) in regulating the cell proliferation and apoptosis of human colorectal cancer cells and explore the possible mechanism.
METHODS
A small interfering RNA (siRNA) targeting AQP5 was used to suppress endogenous AQP5 expression in the human colorectal cancer cell lines COLO 205 and SW480, and the transfection efficiency of AQP5 siRNA was determined using immunofluorescence assay and PCR. The changes in the proliferation of the transfected cells was evaluated with MTT assay, and the cell apoptosis was analyzed using Annexin V-FITC/PI and TUNEL assays; the changes of Bax and Bcl2 expressions in the cells were determined using RTPCR and Western blotting.
RESULTS
Transfection with AQP-5-siRNA resulted in a significant reduction (up to 90%) of AQP-5 expression in COLO 205 and SW480 cells. MTT assay showed that AQP-5-siRNA transfection significantly inhibited the cell proliferation compared NS siRNA transfection (P<0.05). Flow cytometric analysis revealed significantly increased apoptotic rate of cells following AQP-5-siRNA transfection compared with NS?siRNA transfection(P<0.05). Real-time quantitative RT-PCR and Western blotting demonstrated that AQP-5-siRNA transfection significantly increased Bax and Bcl-2 expressions at both mRNA and protein levels in the cells.
CONCLUSION
AQP5-siRNA can promote apoptosis of colorectal cancer cells in vitro possibly in relation to its effects on Bax and Bcl expressions.
目的
研究水通道蛋白5（AQP5）在调控人结肠癌细胞增殖和凋亡中的作用，并探讨其可能机制。
方法
采用靶向AQP5的小干扰RNA（siRNA）抑制人结肠癌细胞系COLO 205和SW480中内源性AQP5的表达，通过免疫荧光分析和PCR检测AQP5 siRNA的转染效率。采用MTT法评估转染细胞的增殖变化，采用Annexin V-FITC/PI法和TUNEL法分析细胞凋亡情况；采用RT-PCR和Western印迹法检测细胞中Bax和Bcl2表达的变化。
结果
用AQP-5-siRNA转染后，COLO 205和SW480细胞中AQP-5的表达显著降低（高达90%）。MTT法显示，与NS siRNA转染相比，AQP-5-siRNA转染显著抑制细胞增殖（P<0.05）。流式细胞术分析显示，与NS siRNA转染相比，AQP-5-siRNA转染后细胞凋亡率显著增加（P<0.05）。实时定量RT-PCR和Western印迹法表明，AQP-5-siRNA转染显著增加了细胞中Bax和Bcl-2在mRNA和蛋白水平的表达。
结论
AQP5-siRNA可能通过影响Bax和Bcl的表达促进体外结肠癌细胞凋亡。
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