摘要
目的探讨miR-26b在结直肠癌侵袭与转移中的作用。方法提取公共芯片数据库数据,分析miR-26b表达水平与淋巴结转移的关系。选用Caco2和DLD1两种结直肠癌细胞系,应用慢病毒感染的方法,构建miR-26b高表达结直肠癌细胞系。通过Transwell迁移和侵袭实验及划痕愈合实验分析上调miR-26b表达对结直肠癌细胞侵袭和转移能力的影响。通过成球实验分析上调miR-26b表达对结直肠癌细胞干细胞表型的影响。结果芯片检测结果显示,有淋巴结转移的结直肠癌患者肿瘤组织中miR-26b表达水平较无淋巴结转移的患者明显升高[(12.04±0.20)比(11.31±0.19),t=2.646,P = 0.010]。体外实验部分,Transwell实验发现,miR-26b高表达的结直肠癌细胞的侵袭细胞数[(16.40±1.36)比(3.80±0.86),t=7.814,P=0.000]和迁移细胞数[(33.40±2.93)比(8.80±2.40),t=6.505,P=0.000]较miR-26b低表达的细胞系显著增多。划痕愈合实验也证实,miR-26b高表达的结直肠癌细胞迁移速度明显加快。miR-26b高表达的结直肠癌细胞在成球速度及成球密度方面均高于miR-26b低表达的细胞系,实验第10天前者球囊内的肿瘤细胞数量显著多于后者[Caco2细胞系:(168.3±11.7)比(54.2±10.8),t=7.185,P=0.002;DLD1细胞系:(4 076.0±409.8)比(1 613.0±210.1),t=5.349,P=0.006]。结论miR-26b可促进结直肠癌细胞的迁移及侵袭能力,并增强肿瘤细胞的干细胞表型,从而促进结直肠癌的浸润与转移。
Objective To investigate the role of miR-26b in the invasion and metastasis of colorectal cancer. Methods Data of public chip databases were extracted to analyze the relationship between miR-26b expression and lymph node metastasis. Two types of colorectal cancer cell lines, Caco2 and DLD1, were selected, and the miR-26b-high colorectal cancer cell line was constructed using the method of lentivirus infection. The effects of up-regulating miR-26b expression on the invasion and metastasis of eolorectal cancer cells were analyzed by Transwell migration and invasion experiment and wound healing assay. The effect of up-regulating miR-26b expression on stem cell phenotype of colorectal cancer cells was analyzed by sphere-formation assay. Results The microarray detection results showed that the expression of miR-26b in tumor tissues of patients with lymph node metastasis was significantly higher than those without lymph node metastasis [(12.04±0.20) vs. (11.31±0.19), t=2.646, P= 0.010]. In the in vitro experiment section, the Transwell experiment results showed that the number of invasive ceils [(16.40±1.36) vs. (3.80±0.86), t=7.814, P=0.000] and migrating cells [(33.40±2.93) vs. (8.80±2.40), t=6.505, P=0.000] in miR-26b-high colorectal cancer cells was significantly higher as compared to miR-26b-low cells (all P〈0.05). Would healing assay also confirmed that the migration speed of miR-26b-high colorectal cancer cells was significantly accelerated. Both the rate and the density of sphere formation were higher in miR-26b-high colorectal cancer cells than those in miR-26b-low colorectal cancer cells [ Caco2 : ( 168.3±11.7) vs. (54.2±10.8), t=7.185, P=0.002; DLDI:(4 076.0±409.8) vs.(1 613.0±210.1), t=5.349, P=0.006]. Conclusion rniR-26b may promote the invasion and metastasis of eolorectal cancer by accelerating the migration and invasion of eolorectal cancer cells and enhancing the stem cell phenotype of tumor cells.
作者
范德军
戎煜明
邹一丰
张凤
林绪涛
吴小剑
Fan Dejun;Rong Yuming;Zou Yifeng;Zhang Feng;Lin Xutao;Wu Xiaojian(Department of Colorectal Surgery;Department of Rheumatology, The Sixth Affiliated Hospital, Sun Yat-sen University, Guangzhou 510655, China;Department of General Dieases, Sun Yat-sen University Cancer Hospital, Guangzhou 510060, China)
出处
《中华胃肠外科杂志》
CAS
CSCD
北大核心
2018年第7期808-813,共6页
Chinese Journal of Gastrointestinal Surgery
基金
国家自然科学基金(81402412)