Effects of methylation of RASSF10 promoter on proliferation and apoptosis of lung cancer cells

BI Jing; YU Rui; HUANG Bo; XIE Zhaofeng

doi:10.3969/j.issn.1009-0460.2019.09.006

Abstract：Objective To detect the methylation of RASSF10 gene promoter in lung cancer cells, and to explore the effect of 5?aza?dC on the proliferation and apoptosis of lung cancer cells. Methods Promoter methylation of RASSF10 gene in human lung cancer cell lines (A549, NCI?H157, SPCA?1, NCI?H460, NCI?H446, QG56) was detected by bisulfite modification combined with restriction endonuclease assay (COBRA). Methylation?positive cell lines were treated with 0,5,10 μmol／L 5?aza?dC and RASSF10 ex?pression was detected by reverse transcription polymerase chain reaction (RT?PCR). MTT assay was used to detect the proliferation ac?tivity of cells, and flow cytometry was used to detect the apoptosis of cells. Results COBRA method detected that SPCA?1 was com?pletely methylated, H446, H157 and H460 were partially methylated, but H157 and H460 were weakly methylated. After 5?aza?dC treatment, RASSF10 mRNA levels in SPCA?1 and H446 cells were significantly up?regulated in a concentration?dependent manner (P＜0. 05). 5?aza?dC could significantly inhibit the proliferation of SPCA?1 and H446 cells in a concentration?and time?dependent manner. The apoptotic rates of SPCA?1 cells in 0 μmol／L 5?aza?dC group,5 μmol／L 5?aza?dC group and 10 μmol／L 5?aza?dC group were (0. 23±0. 10)%, (15. 09±0. 26)% and (34. 9±0. 32)%. The apoptotic rates of H446 cells were (0. 77±0. 10)%, (14. 82±0. 23)%and (35. 4±0. 30)%. The differences were statistically significant (P＜0. 05). Conclusion RASSF10 promoter methylation occurs in human lung cancer cell lines. 5?aza?dC can effectively restore the expression of RASSF10, inhibit the proliferation of lung cancer cells and induce apoptosis.