微小RNA-218在胆管癌中的表达及其对胆管癌细胞凋亡的影响

Expression of microRNA -218 in cho!angiocarcinoma and its effect on the apoptosis of cholangio-carcinoma cells

目的检测胆管癌中微小RNA（miRNA，miR）-218的表达水平及观察其对胆管癌细胞凋亡的影响。方法培养两种胆管癌细胞株（CCLP1）和（QBC939）和非肿瘤胆管细胞株（H69），采用实时定量聚合酶链反应（Real-timePCR）检测miR-218，分为CCLPl、QBC939、H69组，每组随机取3个结果，比较其在胆管癌细胞株与非肿瘤胆管细胞株中的差异。合成miR-218mimics（miR．218激动剂），将miR-218minics及阴性对照转染CCLPl和QBC939细胞。设立miR-218组、阴性对照组及空白对照组，分别利用流式细胞术检测细胞凋亡情况，每组随机取3个结果。结果miR-218在胆管癌细胞株CCLPl、QBC939及非肿瘤胆管细胞株中的相对定量分别为：1．000（以CCLP1为对照）、0．602±0．108、25．199±5．128，miR-218在胆管癌细胞株CCLP1、QBC939中的表达显著低于非肿瘤胆管细胞株（H69）（P=0．000）。CCLPI细胞株中miR-218组细胞凋亡率为0．121±0．010，阴性对照组细胞凋亡率为0．048±0．021，空白对照组细胞凋亡率为0．076±0．017；QBC939细胞株中miR-218组细胞凋亡率为0．118±0。039，阴性对照组细胞凋亡率为0．017±0．036，空白对照组细胞凋亡率为0．048±0．161。miR-218组较阴性对照组及空白对照组细胞凋亡率显著增加，miR-218组与空白对照组及阴性对照组之间的差异有统计学意义（CCLP-1细胞株和QBC939细胞株其P值分别为0．005、0．006）。结论miR-218在胆管癌的发生发展发挥了抑癌因子的作用，并且增加了胆管癌细胞的凋亡。

Objective To detect the expression of miR -218 in cholangiocarcinoma and to study the effect of it on the apoptosis of bile duct cancer cells. Methods Two kinds of cholangiocareinoma cell lines （CCLP1 and QBC939 ） and non - tumor cholangiocarcinoma cell line （t169） were cultured. MiR-218 was detected by real- time quantitative polymerase chain reaction （Real -time PCR） and CCLP1 group, QBC939 group and H69 group were set up. Three results were randomly selected from each group to compare the differences between them. MiR- 218 mimics （miR- 218 agonist） were synthesized and transfected into CCLP1 and QBC939 ceils. MiR- 218 group, negative control group and blank control group were set up. Apoptosis was detected by flow cytometry. Three results were randomly selected from each group. Results The relative quantities of miR - 218 in cholangioearcinoma cell lines CCLP1,QBC939 and non - tumor cholangiocarcinoma cell line （ H69 ） were 1. 000 （ CCLP - 1 control ） , 0. 602 ± 0. 108,25. 199 ± 5. 128, respectively. The expression of miR -218 in cholangiocarcinoma cell lines CCLP1 and QBC939 was significantly lower than that in non - tumor cholangiocarcinoma cell line （ H69 ） （ P = 0. 000）. The apoptosis rate of miR -218 cells in CCLP1 cell line was 0. 121 ±0. 010 and 0. 048 ±0. 021 in negative control group,0. 076 ± 0. 017 in blank control group,0. 118 ± 0. 039 in QBC939 cell line, 0. 017 ± 0. 036 in negative control group and 0. 048 ± 0. 161 in blank control group. The apoptosis rate in miR- 218 group was significantly higher than that in negative control group and blank control group, and the difference between miR- 218 group and blank control group or negative control group was statistically significant （P values for CCLP - 1 cell line and QBC939 cell line were 0. 005 and 0. 006 respectively）. Conclusion MiR -218 plays the role of tumor suppressor factor in the development of cholangiocarcinoma, and increases the apoptosis of bile duct cancer cells.