MiR-885对结直肠癌细胞奥沙利铂化疗敏感性的影响

Effect of MiR-885 on Sensitivity to Oxaliplatin in Colorectal Cancer Cells

[目的]探究miR-885对结直肠癌细胞奥沙利铂化疗敏感性的影响。[方法]利用qRT-PCR验证在耐药和敏感结直肠癌蜡块组织中miR-885的相对表达量;运用质粒转染方法沉默结直肠癌细胞系HCT116中miR-885基因,按照miR-885表达水平分为2组:miR-NC和miR-885inhibitor组,并且利用qRT-PCR验证转染效率。将细胞分成4组,分别是阴性对照组(NC)、阴性对照加奥沙利铂组(L-OHP)、miR-NC加奥沙利铂组(L-OHP/miR-NC)和miR-885 inhibitor加奥沙利铂组(L-OHP/miR-885 inhibitor)。利用CCK-8法检测HCT116细胞活性;Transwell检测HCT116细胞的侵袭能力;流式细胞术检测HCT116细胞的细胞周期。[结果]相比于癌旁正常组织(1.31±0.36),耐药组织(2.75±0.82)中miR-885的表达量增加,而敏感组织(0.31±0.14)中miR-885的表达量降低。在miR-885组中miR-885表达水平(0.02±0.01)相对于miRNC组(0.80±0.19)呈显著降低(P<0.05)。与L-OHP/miR-NC组(0.77±0.14)相比,L-OHP/miR-885 inhibitor组(0.48±0.23)细胞活性降低(P<0.05)。与L-OHP/miR-NC组(169±25)相比,L-OHP/miR-885 inhibitor组(28±9)细胞的侵袭数量减少(P<0.05)。与NC组G_0/G_1期(42.27±3.33)相比,L-OHP组(3.33±0.41)细胞比例降低(P<0.001),与NC组S期(32.19±2.29)相比,L-OHP组(53.29±3.04)细胞比例增多(P<0.05),与NC组(16.32±2.29)G_2/M期相比,L-OHP组(4.83±0.76)细胞比例减少(P<0.05)。同时,与L-OHP/miR-NC组(5.07±0.31)G_0/G_1期相比,L-OHP/miR-885 inhibitor组(45.59±1.95)细胞比例增加(P<0.001),与L-OHP/miR-NC组(42.71±2.58)S期相比,L-OHP/miR-885 inhibitor组(12.72±1.60)细胞比例降低(P<0.001),与L-OHP/miR-NC组(18.19±1.30)G_2/M期相比,L-OHP/miR-885 inhibitor组(2.55±0.51)细胞比例减少(P<0.01)。[结论]沉默miR-885能够增强结直肠癌细胞HCT116对奥沙利铂化疗敏感性,miR-885可能成为结直肠癌治疗的新靶点。

[Purpose]To investigate the effect of miR-885 on chemotherapy sensitivity of colorectal cancer cells.[Methods]The relative expression of miR-885 was detected with qRT-PCR in drug sensitive and resistant colorectal cancer tissues.The human colon cancer HCT116 cells were transfected with miR-885 inhibitor(miR-885 inhibitor)or blank plasmid(miR-NC).Then the cells were divided into four groups:negative control group(NC),negative control group plus oxaliplatin(LOHP),miR-NC plus oxaliplatin group(L-OHP/miR-NC)and miR-885 inhibitor plus oxaliplatin group(L-OHP/miR-885).CCK-8 method was used to detect the viability of HCT116 cells;Transwell method was used to detect the cell invasiveness;flow cytometry was used to detect the cell cycle.[Results]The expression of miR-885 in drug-resistant tissues(2.75±0.82)was higher than that in sensitive tissues(0.31±0.14).The expression level of miR-885 in miR-885 group(0.02±0.01)was significantly lower than that in miR-NC group(0.80±0.19)(P<0.05).The results of CCK-8 showed that compared with L-OHP/miR-NC group(0.77±0.14),the cell viability of L-OHP/miR-885 inhibitor group(0.48±0.23)decreased(P<0.05).Compared with L-OHP/miR-NC group(169±25),the number of migrant cells in L-OHP/miR-885 inhibitor group(28±9)decreased(P<0.05).Compared with NC group(42.27±3.33)in G0/G1 phase,the cell proportion of L-OHP group(3.33±0.41)decreased(P<0.001),compared with NC group(32.19±2.29)in S phase,the cell proportion of L-OHP group(53.29±3.04)increased(P<0.05),compared with NC group(16.32±2.29)G2/M phase,the cell proportion of L-OHP group(4.83±0.76)decreased(P<0.05).Compared with L-OHP/miR-NC group(5.07±0.31)in G0/G1 phase,the cell proportion of L-OHP/miR-885 inhibitor group(45.59±1.95)increased(P<0.001),compared with L-OHP/miR-NC group(42.71±2.58)in S phase,the cell proportion of L-OHP/miR-885 inhibitor group(12.72±1.60)decreased(P<0.001),compared with the G2/M phase of L-OHP/miR-NC group(18.19±1.30),the cell proportion of L-OHP/miR-885 inhibitor group(2.55±0.51)decreased(P<0.01).[Conclusion]MiR-885 silencing can enhance the chemosensitivity of colon cancer HCT116 cells to oxaliplatin,indicating that miR-885 might be a novel target for colorectal cancer treatment.