非编码环状RNA circ_0024597促进大肠癌细胞增殖侵袭研究  

Non-coding circular RNA circ_0024597 in promoting proliferation and invasion of colorectal cancer cells

作　　者：李健[1] 胡均 LI Jian;HU Jun(Tianjin University Hospital,Tianjin 300072,China;Department of Colorectal Cancer Surgery,National Clinical Research Center for Cancer,Key Laboratory of Cancer Prevention and Therapy of Tianjin,Tianjin's Clinical Research Center for Cancer,Tianjin Medical University Cancer Institute and Hospital,Tianjin 300060,China)

机构地区：[1]天津大学医院,天津300072 [2]天津医科大学肿瘤医院结直肠肿瘤科,国家肿瘤临床医学研究中心,天津市肿瘤防治重点实验室,天津市恶性肿瘤临床医学研究中心,天津300060

出　　处：《生物医学工程与临床》2020年第3期317-321,共5页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(81101870);国家临床重点专科建设项目(2013-544);天津市卫生健康委员会重点公关项目(16kg127);CSCO-恒瑞肿瘤研究基金资助项目(Y-HR2017-073)

摘　　要：目的探讨非编码环状RNA circ0024597在调控大肠癌细胞增殖方面的作用。方法选择2018年5月至8月天津医科大学肿瘤医院6例经病理诊断证实为大肠癌患者,年龄41~78岁,平均年龄52.4岁(标准差4.65岁);病理诊断均为腺癌,ⅢA期2例,ⅢB期4例。取该临床病例标本和距离癌变边缘> 5 cm处的癌旁组织。通过高通量circRNA芯片,筛选出高表达于大肠癌组织中的非编码环状RNA circ0024597,通过实时定量聚合酶链反应(qRT-PCR)方法在组织层面(33例病理诊断明确腺癌的Ⅲ期大肠癌患者)验证circ0024597在癌组织和癌旁组织的表达情况;构建针对非编码环状RNA circ0024597的质粒,通过转染人大肠癌Colo-320细胞系,上调circ0024597表达(Colo-320-circl-H细胞组),设计针对circ0024597的小干扰RNA,转染Colo-320细胞系,下调非编码环状RNA circ0024597表达(Colo-320-circl-L细胞组);用CCK-8试剂盒检测细胞增殖的变化和Transwell方法检测细胞侵袭情况。结果大肠癌组织中高表达circ0024597,癌旁组织中极少表达,两者间差异有显著统计学意义(ΔCt:26.850±0.384 vs 18.350±0.468。P <0.000 1)。上调非编码环状RNA circ0024597可以促进大肠癌Colo-320细胞系的增殖(培养72 h,约高于Colo-320细胞1.7倍)和侵袭能力(与Colo-320细胞比,侵袭力为2.310±0.573)(P <0.05);下调非编码环状RNA circ0024597表达则抑制大肠癌Colo-320细胞系的增殖(培养72 h,约低于Colo-320细胞0.5倍)和侵袭力(与Colo-320细胞比,侵袭力为0.580±0.482)(P <0.05)。结论非编码环状RNA circ0024597能够促进大肠癌细胞增殖和侵袭,在大肠癌进展过程中起着重要作用。Objective To investigate the effect of non-coding circRNA circ0024597 in regulating proliferation of colorectal cancer cells. Methods From May to August in 2018, 6 patients with colorectal cancer pathologically confirmed, which aged41-78 years old with mean age of 52.4 years old(standard deviation of 4.65 years old) were enrolled. The pathology results were adenocarcinomas, 2 cases in stage ⅢA and 4 in stage ⅢB. The high-throughput circRNA microarray was used to screen out the non-coding RNA circ0024597 from colorectal cancer tissues and adjacent tissues(edge > 5 cm). The expression of circ0024597 was tested by qRT-PCR(33 cases of adenocarcinoma patients with stage Ⅲ colon cancer). The plasmid targeted at circ0024597 and siRNA was constructed by transfection of colorectal cancer Colo-320 cell line to overexpressed(Colo-320-circl-H cell group), and down-regulated(Colo-320-circl-L cell group) expression of circ0024597. CCK-8 kit was used to detect cell proliferation, and Transwell method was used to detect cell invasion situation. Results The expression of circ0024597 was highly expressed in colorectal cancer tissues, but rarely expressed in adjacent tissues, and the difference was statistically significant(P < 0.000 1). Upregulation of non-coding circRNA circ0024597 could promote proliferation and invasion of Colo-320 cell lines in colorectal cancer(P < 0.05). Down-regulation of circ0024597 could inhibited prolifera-tion(culture for 72-hour, about 0.5 times lower than Colo-320 cells) and invasion(0.580 ± 0.482) of Colo-320 cell lines(P <0.05). Conclusion It is demonstrated that non-coding circRNA circ0024597 could promote proliferation and invasion of colorectal cancer cells, which play an important role in progression of colorectal cancer.

关 键 词：环状RNA circ0024597 大肠癌 细胞增殖 细胞侵袭 

分 类 号：Q75[生物学—分子生物学] R735.3[医药卫生—肿瘤]