长链抗分化非编码RNA在胆管癌中的表达及其对胆管癌侵袭转移的影响

Expression of long anti-differentiation noncoding RNA in cholangiocarcinoma and its effect on invasion and metastasis ability of cholangiocarcinoma

目的:观察长链抗分化非编码RNA(lncRNA ANCR)在胆管癌中的表达及其对胆管癌侵袭转移的影响。方法:利用实时定量聚合酶链反应(PCR)检测2018年6月至2019年9月于武汉同济医院治疗的18例胆管癌患者手术切除标本及癌旁组织中lncRNA ANCR表达水平进行检测;利用慢病毒载体转染构建稳定上下调ANCR胆管癌细胞株TFK-1;利用Transwell和划痕实验验证ANCR对胆管癌侵袭迁移能力的影响;利用裸鼠转移模型验证ANCR对胆管癌远处转移能力的影响。统计学分析均应用SPSS 20.0软件进行,两组间统计学差异通过双侧 t检验进行分析。 结果:胆管癌组织中ANCR表达水平相对于癌症组织呈明显高表达,差异具有统计学意义[(6.78±5.06)倍, t=2.702, P<0.05]。上调ANCR可显著增加TFK-1细胞侵袭细胞数[(2 180.00±124.83)个比(952.00±74.47)个, t=16.255, P<0.05]、迁移率[(86.00±3.06)%比(49.00±3.87)%, t=8.416, P<0.01],下调ANCR可显著降低TFK-1细胞侵袭细胞数[(256.00±20.53)个比(952.00±74.47)个, t=-16.789, P<0.05]、迁移率[(29.00±2.58)%比49.00±3.87)%, t=-12.766, P<0.01]。上调ANCR可显著增加TFK-1在裸鼠体内转移灶的形成,下调ANCR可显著降低TFK-1在裸鼠体内转移灶的形成,第5、6周期裸鼠体重[第5周(14.56±0.61) g比(17.33±0.84) g, t=-2.724, P<0.05和(20.62±1.68) g比(17.33±0.84) g, t=3.759, P<0.05;第6周(14.21±0.40) g比(17.98±0.43) g, t=-4.795, P<0.05和(21.48±1.33) g比(17.98±0.43) g, t=8.154, P<0.05]及转移瘤平均直径[第5周(21.00±2.76) mm比(14.80±0.75) mm, t=13.426, P<0.05和(4.40±1.36) mm比(14.80±0.75) mm, t=-7.589, P<0.05;第6周(23.80±3.54) mm比(16.60±1.74) mm, t=7.561, P<0.05和(6.40±2.06) mm比(16.60±1.74) mm, t=-4.491, P<0.05]较对照组差异有统计学意义。 结论:lncRNA ANCR在人胆管癌组织中呈现显著高表达,其可促进胆管癌细胞侵袭、迁移和远处转移能力。

Objective To detect the role of Long anti-differentiation noncoding RNA(lncRNA ANCR)in human cholangiocarcinoma and explore its effect on apoptosis and invasion ability of cholangiocarcinoma.Methods We detected the expression of ANCR in human cholangiocarcinoma tissues,normal bile duct tissues by PCR.And we detected invasion and migration ability of human cholangiocarcinoma cell line TFK-1 by Transwell assay and Wound Healing after ANCR lentivirus transfection.And we detected metastasis ability of TFK-1 by building a BALB/c nude mice metastasis model.Two-tailed Student’s t-test was used to evaluate the differences between two groups.All statistical analyses were performed using SPSS 20.0.Results The expression of ANCR in human cholangiocarcinoma tissues is significantly higher than the normal bile duct tissues[(6.78±5.06)times,t=2.702,P<0.05];The invasion and migration ability of TFK-1 is declined after ANCR-down lentivirus transfection(256.00±20.53 vs.952.00±74.47,t=-16.789,P<0.05).The invasion and migration ability of TFK-1 is improving after ANCR-up lentivirus transfection(2180.00±124.83 vs.952.00±74.47,t=16.255,P<0.05).The metastasis ability of TFK-1 is declined after ANCR-down lentivirus transfection[(29.00±2.58)%vs.(49.00±3.87)%,t=-12.766,P<0.01].The metastasis ability of TFK-1 is improving after ANCR-up lentivirus transfection[(86.00±3.06)%vs.(49.00±3.87)%,t=8.416,P<0.01].ANCR-up cells had formed larger tumors compared with those in control groups[the 5th week,(21.0±2.76)mm vs.(14.8±0.75)mm,t=13.426,P<0.05;the 6th week,(23.8±3.54)mm vs.(16.6±1.74)mm,t=7.561,P<0.05].The weight of ANCR-up group is reduced than those in the negative control group[the 5th week,(14.56±0.61)g vs.(17.33±0.84)g,t=-2.724,P<0.05;the 6th week,(14.21±0.40)g vs.(17.98±0.43)g,t=-4.795,P<0.05]ANCR-down cells had formed smaller tumors compared with those in control groups[the 5th week,(4.40±1.36)mm vs.(14.8±0.75)mm,t=-7.589,P<0.05;the 6th week,(6.40±2.06)mm vs.(16.6±1.74)mm,t=-4.491,P<0.05].The weight of ANCR-down group is increased than those in the negative control group[the 5th week,(20.62±1.68)g vs.(17.33±0.84)g,t=3.759,P<0.05;the 6th week,(21.48±1.33)g vs.(17.98±0.43)g,t=8.154,P<0.05].Conclusion These results indicate that the expression of ANCR is different between cholangiocarcinoma and normal bile duct tissues and the ANCR probably involved in cholangiocarcinoma invasion,migration and metastasis and are the indicator of diagnosis and prognosis of the cholangiocarcinoma.