RESUMEN
Objective To observe the expression of pleckstrin homology like domain family A member 2 (PHLDA2) in hepatocellular carcinoma (HCC) and investigate the effects of PHLDA2 on cell proliferation, migration, invasion and apoptosis. Methods To analyze the expression of PHLDA2 in 369 cases of HCC tissues and 160 cases of adjacent normal tissues and the effect of PHLDA2 expression on overall survival rate of HCC patients by gene expression profiling interactive analysis (GEPIA) online database. Cell proliferation was determined by cell counting kit-8 (CCK-8) and colony formation assay. Invasion and migration were detected by Transwell assay. The percentage of apoptosis was measured by flow cytometry. The levels of Bax and cleaved Caspase-3 were measured by Western blotting. Results The expression of PHLDA2 was upregulated in HCC, and high expression of PHLDA2 reduced the overall survival of HCC patients. Low-expression of PHLDA2 inhibited proliferation, invasion and migration, and increased apoptosis of HCC cells. Conclusion PHLDA2 promotes the occurrence and development and may act as a tumor promoter in HCC.
RESUMEN
Objective To study the effects of PHLDA2 overexpression on radiosensitivity and the underlying mechanisms in human osteosarcoma U2OS cell line.Methods To obtain the subclone,cells were exposed to G418 persistently after transfection of pEGFP-C3-PHLDA2 vector into U2OS cells.Three groups of blank control (U2OS),negative control (U2OS-neo) and transfected group (U2OS-PHLDA2) were used.The expression of PHLDA2 in the subclone cells was determined by Western blot.After exposure to X-ray irradiation,cellular growth activity and survival were detected by CKK-8 assay and colony formation assay,respectively.The cell apoptosis was measured by the Annexin V/PI staining,and the apoptotic protein was analyzed by Western blot.The in-vivo effects of PHLDA2 on irradiation were evaluated by xenografts.Results Compared with U2OS group and U2OS-neogroup,the sabclone cells were successfully obtained by G418 selection,in which the expression of PHLDA2 was upregulated(t =13.73,16.28,P ＜ 0.05).In vitro,PHLDA2 overexpression significantly enhanced the response to radiation in U2OS cells with a reduction of colony survival and proliferation with the increase of doses (t =5.00-8.23,P ＜0.05;t =-2.52--1.26,P ＜ 0.05).In vivo,PHLDA2-upregulated xenografts had more radiosensitivity than control groups with a significant inhibition of tumor growth (t =3.27,2.91,P ＜ 0.05).After 8 Gy irradiation,the apoptosis was significantly increased (t =10.11,9.61,P ＜ 0.05),accompanied with the activation of Caspased-3 in U2OS-PHLDA2 cells,which was presented by upregulation of cleaved Caspase-3 (t =11.26,10.72,P ＜ 0.05).Conclusions Exogenetic expression of PHLDA2 could significantly enhance the radiosensitivity of human osteosarcoma cells,which may be attributed to the activation of Caspase-3 that increases irradiation-induced apoptosis.