长链非编码RNA前列腺癌非编码核糖核酸1在结直肠癌组织中的上调对增殖及细胞周期的影响

摘要

目的 观察长链非编码RNA(IncRNA)前列腺癌非编码核糖核酸1(PRNCR1)在结直肠癌中的表达及其对癌细胞增殖能力的影响,并在体外研究PRNCR1的功能.方法 入组63例经病理确诊为结直肠腺癌患者的癌组织及癌旁组织,实时定量聚合酶链反应(Real-time PCR)法检测PRNCR1的表达水平.设计合成PRNCR1的靶向等位基因特异性寡核苷酸(ASO),通过脂质体介导的转染法转入HT-29细胞,Real-time PCR法检测干扰后HT-29细胞内PRNCR1的转录水平.实验分Scramble转染对照组和PRNCR1-ASO转染实验组:细胞计数试剂盒(CCK4)实验检测PRNCR1低表达对增殖活性的影响;Transwell实验和Matrigel实验检测PRNCR1低表达对结直肠癌侵袭转移的影响.结果 PRNCR1在CRC肿瘤组织中高表达(平均上调10.5倍),并与结直肠癌大小显著相关(P＜0.01).CCK-8实验结果显示:沉默PRNCR1表达后,24、48、72、96 h实验组450 mm吸光度值较对照组显著降低(P＜0.05).结论 PRNCR1是结直肠癌的特异性长链非编码RNA,能促进结直肠癌的增殖.%Objective To investigate the expression of long non-cording RNA (lncRNA) prostate cancer non-coding RNA 1 (PRNCR1) in colorectal cancer (CRC) and its effects on proliferation of CRC cells,and probe the biological function of PRNCR1 in vitro.Methods Real-time quantitative polymerase chain reaction (Real-time PCR) was used to assess whether PRNCR1 is detectable or altered in CRC tissues or cell lines compared to adjacent normal tissues or normal cell lines in a cohort of 63 patients.The PRNCR1 targeted allele-specific oligonucleotide (ASO) was synthesized and transfected into HT-29 cells via the liposome mediation,then Real-time PCR was used to evaluate the PRNCR1 level after ASO-mediated inhibition in HT-29 cells.A Scramble transfection method was used in the control group.The impact of PRNCR1 low-expression on proliferative activity,invasion and metastasis was explored by cell counting kit-8 (CCK-8),Transwell and Matrigel assays,respectively.Results PRNCR1 levels were dramatically increased in both CRC tissues and cell lines compared to controls (10.5 fold on average),and the PRNCR1 expression level was positively correlated with the tumor volume significantly (P ＜ 0.01).Additionally,PRNCR1 was found to influence the proliferation and cell cycle progression of CRC cells.CCK-8 assay manifested that 450 mm absorbance value of treatment group was significantly declined at 24,48,72,and 96 h after silencing PRNCR1 (P ＜ 0.05).Conclusion PRNCR1 is a CRC specific lncRNA,which promotes proliferation of CRC cells.