Effect of SNP rs6983267-related non-coding RNA G/U on the metastatic activity of colon carcinoma cells

CAI Wenchen; HAO Xiaohui; SHEN Yangli; MENG Chenxue; TIAN Jie; GUO Zhiyi

doi:10.3969/j.issn.1002-266X.2016.45.003

Abstract：Objective To investigate the effect of SNP rs6983267 in the 8q24 region on chromosome-related non-cod-ing RNA G/U on the metastatic activity of colon carcinoma cells.Methods We analyzed the non-coding RNA of SNP rs6983267 by bioinformatics.The KM12C and KM12SM cells were collected,and the total RNA was extracted from the cells.Reverse transcription and PCR amplification were performed.After the fragments were recovered,they were digested with Tsp45Ⅰ,and then the digested (digestion group)and non-digested templates (control group)received PCR amplifi-cation analysis.The non-coding RNA G/U expression of SNP rs6983267 in the two groups was detected by restriction endo-nuclease-PCR.pGL3-Gs-LUC (pGL3-Gs-LUC group),pGL3-Ts-LUC (pGL3-Ts-LUC group)and pGL3-Basic-LUC were respectively transfected into KM12C and KM12SM cells.The non-coding RNAs of transcriptional activity in KM12C and KM12SMcells were detected by the dual luciferase reporter gene analysis.Results The ENCODE database showed that non-coding genes existed near the SNP rs6983267 locus in the 8q24 region on chromosome.In the KM12C cells,the non-coding RNA G/U ratios of the SNP rs6983267 in the digestion group and control group were 0.71 ±0.09 and 1.43 ±0.11, and the ratios were 0.80 ±0.10 and 5.60 ±1.30 in the KM12SMcells.Compared with the control group,the non-coding RNA G/U ratio decreased in the KM12C cells and increased in the KM12SMcells (all P <0.05).In KM12C cells,the relative expression activity of the G promoter and T promoter on the rs6983267 locus was 0.81 ±0.14 in the pGL3-Gs-LUC group,and 0.82 ±0.25 in the pGL3-Ts-LUC group,and in the KM12SMcells they were 1.00 ±0.26 and 1.85 ±0.58, respectively.In KM12SMcells,statistically significant difference was found between these two groups (P <0.05),while in the KM12C cells,no statistically significant difference was found (P >0.05 ).Conclusion The G/U ratio of SNP rs6983267-related non-coding RNA may be associated with colon cancer metastasis.