长基因间非蛋白质编码RNA525在结直肠癌组织的表达及其对结直肠癌细胞增殖、侵袭的影响

目的观察长基因间非蛋白质编码RNA525(LIN00525)在结直肠癌中的表达及其对结直肠癌细胞体外增殖、侵袭的影响。方法收集2018年8月至2018年12月于复旦大学附属肿瘤医院大肠外科诊治患者的50对结肠癌及癌旁组织,通过实时定量聚合酶链式反应(Real time-PCR)检测50对样本中LINC00525的表达并分析其表达水平与临床病理参数的相关性;将LINC00525的过表达质粒转染SW480细胞,敲减质粒转染HCT-8细胞,通过细胞计数试剂盒(CCK-8)增殖实验、Transwell侵袭实验观察LINC00525对SW480及HCT-8细胞体外增殖、侵袭力的影响;通过蛋白质印迹法(Western blot)实验检测增殖相关分子增殖细胞核抗原(PCNA)及侵袭相关分子基质金属蛋白酶(MMP)-2的表达探索LINC00525调控结直肠癌细胞增殖、侵袭的分子机制;应用SPSS 20.0统计软件分析,临床资料分析使用χ²检验,两样本均数间的比较采用Student’t检验。结果50对结直肠癌组织中LINC00525的表达显著高于癌旁组织(0.721±0.028比0.204±0.016,t=15.860,P<0.01),差异有统计学意义;LINC00525的表达水平与结直肠癌淋巴转移呈正相关(χ²=10.092,P<0.01),差异有统计学意义;CCK-8实验结果显示LINC00525高表达组的吸光度(A)值显著高于对照组(2.163±0.101比1.250±0.035,t=8.525,P<0.01),差异有统计学意义;敲减LINC00525组的A值显著低于对照组(0.717±0.102比1.483±0.103,t=5.305,P<0.01),差异有统计学意义;Transwell侵袭实验结果提示过表达LINC00525组的侵袭细胞数量显著高于对照组(45.670±6.936比87.670±4.910,t=4.942,P<0.01),差异有统计学意义;敲减LINC00525组的细胞侵袭数量显著低于对照组(81.670±8.570比43.330±5.364,t=3.791,P<0.05),差异有统计学意义;Western blot显示过表达LINC00525组PCNA及MMP-2的的表达量分别高于对照组(1.72比1.00,t=5.971,P<0.01;1.58比1.00,t=7.197,P<0.01),差异有统计学意义;敲减LINC00525组PCNA及MMP-2的表达量分别低于对照组(0.39比1.00,t=7.739,P<0.01;0.33比1.00,t=12.170,P<0.01),差异有统计学意义。结论LINC00525在结直肠癌组织中高表达,LINC00525通过调控PCNA及MMP-2的表达影响结直肠癌细胞增殖和侵袭。

Expression of long intergenic non-protein coding RNA 525 in colorectal cancer tissues and its effect on the proliferation and invasion of colorectal cancer cells

Objective To study the expression of long intergenic non-protein coding RNA 525(LIN00525)in colorectal cancer(CRC)and its effect on the proliferation and invasion of CRC cells in vitro.Methods Totally 50 pairs of colon cancer and adjacent tissues were collected from patients diagnosed and treated in the Department of Colorectal Surgery,Fudan University Shanghai Cancer Center during August 2018 to December 2018.Real-time quantitative polymerase chain reaction(RT-PCR)was used to detect the expression of LINC00525 in 50 pairs of colon cancer and adjacent tissues.The correlation between the expression level of LINC00525 and clinicopathological parameters was analyzed.The overexpression plasmid and knockdown plasmid of LINC00525 were transfected into SW480 and HCT-8 cells,respectively.The cell counting kit-8(CCK-8)proliferation assay and Transwell invasion assay were used to observe the in vitro proliferation and invasion of LINC00525 on intestinal cancer cell lines.The expression of proliferation-related protein proliferating cell nuclear antigen(PCNA)and invasion-related protein matrix metalloproteinase(MMP)-2 was detected by Western blotting to explore the molecular mechanism of LINC00525 regulating the proliferation and invasion of CRC cells.The SPSS 20.0 statistical software was used for analysis,the clinical data analysis was performed by theχ² test,and the comparison between the two sample means was performed by the Student′s t test.Results The expression of LINC00525 in 50 pairs of CRC tissues was significantly higher than that in adjacent tissues(0.721±0.028 vs.0.204±0.016,t=15.86,P<0.01).The expression level of LINC00525 was positively correlated with lymphatic metastasis of CRC(χ²=10.092,P<0.01).CCK-8 assay showed that the absorbance value in LINC00525 high expression group was significantly higher than that in control group(2.163±0.101 vs.1.25±0.035,t=8.525,P<0.01).The absorbance value in the knock-down LINC00525 group was significantly lower than that in the control group(0.717±0.102 vs.1.483±0.103,t=5.305,P<0.01).Transwell invasion experiment showed that the number of invasion cells in the overexpression LINC00525 group was significantly greater than that in the control group(45.670±6.936 vs.87.670±4.910,t=4.942,P<0.01).The number of invasive cells in the LINC00525 knockdown group was significantly less than that in the control group(81.670±8.570 vs.43.330±5.364,t=3.791,P<0.05).Western blotting showed that the expression levels of PCNA and MMP-2 in the LINC00525 overexpression group were significantly higher than those in the control group(1.72 vs.1.00,t=5.971,P<0.01;1.58 vs.1.00,t=7.197,P<0.01).The expression levels of PCNA and MMP-2 in the LINC00525 knock-down group were significantly lower than those in the control group(0.39 vs.1.00,t=7.739,P<0.01;0.33 vs.1.00,t=12.170,P<0.01).Conclusion LINC00525 is highly expressed in CRC tissues.LINC00525 affects the proliferation and invasion of CRC cells by regulating the expression of PCNA and MMP-2.