Rs217727 polymorphism in H19 promotes cell apoptosis by regulating the expressions of H19 and the activation of its downstream signaling pathway.

BACKGROUND

The objective of the current study was to explore the role of H19 rs217727 polymorphism in the control of hepatocellular carcinoma (HCC).

METHOD: The Student's t test, Cox regression, and Kaplan-Meier analyses were used to clarify whether the H19 rs217727 polymorphism played an important role in the development of HCC. Real-time polymerase chain reaction (PCR) and western-blot analysis were carried out to measure the levels of H19, microRNA (miR)-675, FAS-associated death domain (FADD), caspase-8, and caspase-3 among H19 CC, CT, and TT groups, as well as in cells transfected with H19/si-H19, or miR-675 mimic/inhibitor. The MTT assay, colony formation assay, and flow cytometry assay were performed to detect the effect of H19/miR-675 on cell viability, cell colony formation, and cell apoptosis.

RESULT: T allele of H19 rs217727 polymorphism apparently increased the survival rate of patients with HCC. Meanwhile, H19 enhanced miR-675 expression but reduced the mRNA and protein levels of FADD, caspase-3, and caspase-8. The T allele of H19 rs217727 polymorphism apparently increased the apoptotic rate of HCC cells. Furthermore, FADD was a virtual target gene of miR-675 with a potential "hit" located in the 3'-untranslated region (UTR) of FADD, whereas H19 inhibited FADD expression via increasing the expression of miR-675. Moreover, H19 upregulated the expression of miR-675 whereas reducing the expression of FADD, caspase-3, and caspase-8. Finally, H19 and miR-675 promoted cell proliferation and cell colony formation but repressed cell apoptosis.

CONCLUSION

In summary, the above findings demonstrated that the polymorphism of rs217727 in H19 was associated with HCC via the H19/miR-675/FADD/caspase-8/caspase-3/apoptosis signaling pathway.