摘要
目的探讨长链非编码RNA(LncRNA)RAB11B-AS1对耐吉西他滨结直肠癌细胞增殖、凋亡的影响及分子机制。方法选取温州市中西医结合医院2017年1月至2019年1月37例结直肠癌患者的癌组织和癌旁组织。采用逐步增加培养基中吉西他滨的浓度,建立耐吉西他滨的结直肠癌细胞Caco-2-Gem;将其分为si-NC组、si-RAB11B-AS1组、si-RAB11B-AS1+anti-miR-NC组、si-RAB11B-AS1+anti-miR-486-5p组。以实时荧光定量PCR(RT-qPCR)对miR-486-5p及RAB11B-AS1的表达情况进行检测;分别用0、1、5、10、50、100、500μmol/L吉西他滨处理Caco-2和Caco-2-Gem细胞,以四甲基偶氮唑盐比色法(MTT)对细胞存活率进行检测;以流式细胞术对细胞周期、凋亡情况进行检测。结果和Caco-2细胞、癌旁组织相比,结直肠癌组织和Caco-2-Gem细胞中RAB11B-AS1表达水平升高(P<0.05),miR-486-5p表达水平降低(P<0.05)。与0μmol/L组相比,1~100μmol/L吉西他滨处理后Caco-2细胞存活率均显著降低(P<0.05);1~10μmol/L吉西他滨处理后Caco-2-Gem细胞存活率无明显变化,Caco-2-Gem细胞存活率经吉西他滨(50~500μmol/L)处理后降低(P<0.05)。抑制表达LncRNA RAB11B-AS1后,Caco-2-Gem细胞存活率降低,凋亡率升高,S期细胞占比降低,G0-G1期细胞占比升高(P<0.05)。在抑制LncRNA RAB11B-AS1表达的基础上下调miR-486-5p表达后,Caco-2-Gem细胞存活率升高,细胞凋亡率降低,S期细胞占比升高,G0-G1期细胞占比降低。结论抑制表达lncRNA RAB11B-AS1可能对miR-486-5p具有负调控作用,从而在促进对吉西他滨有耐药能力结直肠癌细胞凋亡的同时抑制其增殖,提高吉西他滨对结直肠癌细胞的生物效能。
ObjectiveTo explore the effect and molecular mechanism of long non-coding RNA(LncRNA)RAB11B-AS1 on the proliferation and apoptosis of colorectal cancer cells.MethodsCancerous tissues and para-cancerous tissues of 37 patients with colorectal cancer from Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine from Jan.2017 to Jan.2019 were selected.By gradually increasing the concentration of gemcitabine in the medium,the colorectal cancer cell Caco-2-Gem resistant to gemcitabine was established and divided into si-NC group,si-RAB11B-AS1 group,si-RAB11B-AS1+anti-miR-NC group,and si-RAB11B-AS1+anti-miR-486-5p group.The expression of miR-486-5p and RAB11B-AS1 was detected by real-time quantitative PCR(RT-qPCR);Caco-2 and Caco-2-Gem cells were treated with 0,1,5,10,50,100,and 500μmol/L gemcitabine,respectively;Cell viability was measured by MTT assay;The cell cycle and apoptosis were detected by flow cytometry.ResultsCompared with Caco-2 cells and adjacent tissues,the expressions of RAB11B-AS1 was increased in colorectal cancer tissues and Caco-2-Gem cells(P<0.05),while the expression of miR-486-5p was decreased(P<0.05).Compared with the 0μmol/L group,the survival rate of Caco-2 cells after 1-100μmol/L gemcitabine treatment was significantly reduced(P<0.05);the survival rate of Caco-2-Gem cells had no significant change after 1-10μmol/L gemcitabine treatment,and the survival rate of Caco-2-Gem cells was decreased after treatment with Gissi(50-500μmol/L)(P<0.05).After inhibiting the expression of LncRNA RAB11B-AS1,the survival rate of Caco-2-Gem cells decreased,the apoptosis rate increased,the proportion of cells in S phase decreased,and the proportion of cells in G0-G1 phase increased.After inhibiting the expression of LncRNA RAB11B-AS1 and down-regulating the expression of miR-486-5p,the survival rate of Caco-2-Gem cells was increased,the apoptosis rate was decreased,the proportion of cells in S phase was increased,and the proportion of cells in G0-G1 phase was decreased(P<0.05).ConclusionsInhibition of the expression of the lncRNA RAB11B-AS1 may have a negative regulatory effect on miR-486-5p,thereby inhibiting proliferation while promoting apoptosis in colorectal cancer cells resistant to Gissi,thus improving the biological efficacy of Gissi on colorectal cancer cells.
作者
袁玉青
张煜程
胡业晓
郑艳艳
Yuan Yuqing;Zhang Yucheng;Hu Yexiao;Zheng Yanyan(Department of Proctology,Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine,Wenzhou 325000,China;Department of General Surgery,Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine,Wenzhou 325000,China)
出处
《中华内分泌外科杂志（中英文）》
CAS
2024年第6期875-880,共6页
Chinese Journal of Endocrine Surgery
基金
温州市科技计划项目(Y20170444)。
