摘要目的 了解巨噬细胞加帽蛋白(CapG)在结直肠癌组织中的表达,探讨其对结直肠癌细胞增殖、迁移能力的影响.方法 收集2015年9月10日至2016年3月2日行结直肠癌手术治疗的84例患者的临床资料和组织标本,其中结直肠癌组织标本84份,结直肠癌旁正常组织(对照)19份.采用免疫组织化学法检测结直肠癌组织和癌旁正常组织中CapG的表达情况.将结直肠癌HCT116细胞分为阴性对照组和小干扰RNA组,分别转染阴性对照序列和靶向CapG的序列.采用实时荧光定量PCR和蛋白质印迹法检测干扰效果.采用MTT法、细胞划痕实验和Transwell细胞迁移实验检测干扰CapG后对细胞增殖和迁移的影响.统计学分析分别采用t检验、Wilcoxon秩和检验和Kruskal-Wallis H检验.结果 CapG在结直肠癌组织的阳性表达率为95％(80/84),高于癌旁正常组织(5/19),差异有统计学意义(U=-6.123,P=0.023).CapG在结直肠癌患者不同发生部位、有无淋巴结转移、不同分化程度和不同临床分期中的表达差异均有统计学意义(H=10.683,U=2.668,H=12.182,U=-3.247;P均＜0.05).阴性对照组和小干扰RNA组的HCT116细胞增殖速度差异均无统计学意义(P均＞0.05).小干扰RNA组HCT116细胞株的划痕愈合率为(27.40±1.58)％,低于阴性对照组的(64.40±1.25)％,差异有统计学意义(t=31.790,P＜0.01).小干扰RNA组HCT116细胞株的穿膜细胞数为57.0±2.0,少于阴性对照组的110.0±6.7,差异有统计学意义(t=13.290,P＜0.01).结论 结直肠癌组织中CapG表达增高,其高表达可能与结直肠癌的转移密切相关.

abstractsObjective To explore the expression of maerophage capping protein (CapG) in colorectal carcinoma tissues,and to investigate its effects on proliferation and migration of colorectal carcinoma cells.Methods From September 10th,2015 to March 2nd,2016,the clinical data and tissues specimen of 84 patients with colorectal operation were collected.Eighty-four specimens were colorectal cancer tissue and nineteen specimens were adjacent normal tissue (control).The expression of CapG in colorectal cancer tissue and adjacent normal tissue were detected by immunohistochemistry.The colorectal carcinoma HCT116 cells were divided into negative control group and small interfering RNA (siRNA) group,which were transfected with negative control siRNA and CapG siRNA,respectively.The interfering effects were identified by real-time polymerase chain reaction (qRT-PCR) and Western blotting.The effects of CapG interfering on cell proliferation and migration were determined by 3-(4,5-dimethyl-2-tbiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method,wound healing assay and Transwell migration assay.Student t test,Wilcoxon Mann-Whitney test,and Kruskal-Wallis H test were performed for statistical analysis.Results The positive expression rate of CapG in colorectal cancer tissue was 95％ (80/84),which was higher than that of adjacent normal tissue (5/19),and the difference was statistically significant (U=-6.123,P =0.023).The differences of CapG expression in patients with colorectal cancer at different location,with or without lymph node metastasis,different differentiation degree and different clinical stage were all statistically significant (H=10.683,U=2.668,H=12.182,U=-3.247,respectively;all P＜0.05).There was no statistically significant differnce in the proliferation of HCT116 cells between negative control group and siRNA group (all P＞0.05).The rate of wound healing of HCT116 cells in siRNA group was (27.40 ± 1.58)％,which was lower than that of negative control group((64.40±1.25)％),and the difference was statistically significant(t=31.790,P＜0.01).The number of migrated cells through the micropore membrane of siRNA group was 57.0± 2.0,which was less than that of negative control group(110.0± 6.7),and the difference was statistically significant (t=13.290,P＜0.01).Conclusion The expression of CapG in colorectal carcinoma tissue increases,which might be closely associated with the metastasis of colorectal cancer.