长链非编码RNA LINC01296对胃癌细胞增殖的影响及其机制

Effect and mechanism of long non-coding RNA LINC01296 on proliferation of gastric cancer cells

目的观察长链非编码RNA LINC01296对胃癌细胞增殖的影响及其机制。方法收集人胃癌细胞系SGC-7901、BGC-823及人正常胃黏膜上皮细胞GES1,采用实时荧光定量PCR法检测细胞LINC01296表达。将SGC-7901、BGC-823细胞各分为两部分,分别加入小干扰RNA(siRNA)、阴性对照RNA(siNC)进行转染,转染24 h。收集细胞,采用CCK-8法检测波长450 nm处的光密度(OD)值,克隆形成实验计数克隆形成数,胞质胞核分离实验观察LINC01296在胃癌细胞中的分布,RNA结合蛋白免疫共沉淀实验观察LINC01296与EZH2、SUZ12的相互作用。结果SGC-7901、BGC-823及GES1细胞LINC01296相对表达量分别为4.04±1.64、5.33±1.32、1.41±0.01,SGC-7901、BGC-823细胞LINC01296相对表达量均高于GES1细胞(P均<0.05)。转染siRNA、siNC的SGC-7901细胞OD值分别为1.20±0.03、1.38±0.02,克隆形成数分别为(112.56±5.22)、(256.14±6.42)个,二者比较P均<0.05;转染siRNA、siNC的BGC-823细胞OD值分别为1.50±0.04、1.73±0.22,克隆形成个数分别为(98.21±5.56)、(203.76±5.31)个,二者比较P均<0.05。LINC01296在SGC-7901、BGC-823细胞核中的相对表达比例均高于细胞质(P均<0.05)。RNA结合蛋白免疫共沉淀实验结果显示,LINC01296可以与EZH2、SUZ12结合。结论LINC01296可能通过与EZH2、SUZ12结合而促进胃癌细胞增殖。

Objective To observe the effect and mechanism of long non-coding RNA(LINC01296)on the proliferation of gastric cancer cells GES1.Methods We collected the human gastric cancer cell lines SGC7901,BGC823,and normal gastric epithelial cells GES1.The expression of LINC01296 in the above cells was detected by real-time quantitative PCR.SGC-7901 and BGC-823 cells were divided into two parts,which were transfected with small interfering RNA(siRNA)and control siRNA(siNC),respectively,after 24 hours,the cells were collected.CCK-8 assay was used to detect the OD value at a wavelength of 450 nm(OD value).The clone formation assay was used to detect the number of clone formation.The subcellular fractionation assay was used to observe the distribution of LINC01296 in gastric cancer cells.RNA immunoprecipitation(RIP)experiment was used to observe the interaction of LINC01296 with EZH2 and SUZ12.Results The expression levels of LINC01296 in SGC7901,BGC823 and GES1 cells was 4.039±1.64,5.33±1.32,and 1.41±0.01,respectively,which indicated that the expression level of LINC01296 in SGC7901 and BGC823 cells was higher than that of GES1 cells.The OD value of SGC7901 cells transfected with siRNA and siNC were 1.20±0.03 and 1.38±0.02,respectively,and the number of clone formations were 112.56±5.22 and 256.14±6.42,respectively(both P<0.05);The OD values of BGC823 cells transfected with siRNA and siNC were 1.50±0.04 and 1.73±0.22,respectively.The number of clones formations was 98.21±5.56 and 203.76±5.31(both P<0.05).The relative expression of LINC01296 in the nuclei of SGC7901 and BGC823 cells was higher than that of cytoplasm(both P<0.05).RIP experiments showed that LINC01296 can be combined with EZH2 and SUZ12.Conclusion LINC01296 is highly expressed in the gastric cancer and could promote the proliferation of gastric cancer cells through combining with EZH2 and SUZ12.