Effects of FAM96B on the proliferation, apoptosis, invasion and metastasis of hepatocellular carcinoma HepG2 cells

Abstract

Objective To investigate the effects of family with sequence similarity 96 member B (FAM96B) on the proliferation, apoptosis, invasion and metastasis of hepatocellular carcinoma (HCC) HepG2 cells. Methods FAM96B mRNA and protein were examined respectively by real-time fluorogenic quantitative polymerase chain reaction (Real-time PCR) and Western blotting in L02 and HepG2 cells. Moreover, the eukaryotic expression vector of FAM96B was constructed and transfected into HepG2 cells. The cell proliferation of each group at 0, 24, 48, 72 and 96 h was tested by methyl thiazol tetrazolium (MTT). Then the apoptosis, invasion and metastasis were tested respectively by Flow cytometry, wounding healing assay and Transwell. Meanwhile, the subcutaneous tumorigenic model of nude mice were established by 10 male and aged 4-5 weeks BALB/c-nu/nu mice. The model mice were randomized into control group and FAM96B group, 5 mice in each group. And which were injected with 0.3 ml 1×1010 pcDNA3.1-HepG2 cells and pcDNA3.1-FAM96B HepG2 cells. The weight and volume of tumors were measured every three days. All the mice were executed on the 30th day. Statistical analysis was performed using SPSS 21.0. The measurement data were expressed as mean±standard deviation (Mean±SD). The mean comparison between the two groups was performed by t test. Results The FAM96B mRNA expression was decreased in HepG2 cells than in L02 cells (t=25.343, P<0.01). Meanwhile, The FAM96B protein expression was decreased in HepG2 cells than in L02 cells. Compared with control group, the proliferative ability of FAM96B group was significantly reduced (P<0.01). The apoptosis rate in FAM96B group was increased in FAM96B group than in control group [(8.61±1.05)% vs. (47.34±4.82)%, t=23.544, P<0.01]. Compared with control group, the migration ability of FAM96B group was obviously reduced [(222.33±32.78) μm vs. (284.13±46.18) μm, t=4.227, P<0.01], and the invasion ability was obviously reduced [(45.47±4.05) vs. (101.13±5.54), t=31.413, P<0.01]. Moreover, in the subcutaneous tumorigenic model of nude mice, the tumour volume and weight were all decreased in FAM96B group than in control group (tvolume=0.610, tweight=7.186, P all<0.05). Conclusion FAM96B in hepatocellular carcinoma HepG2 cells is low expression, up-regulated its expression levels can inhibit the proliferation, invasion, metastasis and induce apoptosis of HepG2 cells. And up-regulated FAM96B expression levels can inhibit growth of tumors in the subcutaneous tumorigenic model of nude mice. Key words: Family with sequence similarity 96 member B; Hepatocellular carcinoma; HepG2 cells; Proliferation; Apoptosis; Invasion and metastasis