Effect Of MiR-21 On The Proliferation Of Hepatocytes Induced By Acrylamide And Its Combined Alcohol And The Intervention Of Curcumin

Objective: Acrylamide, a proven rodent carcinogen, is a common chemical that widely exists in daily life. Acrylamide can be metabolized into glycidamide mainly by cytochrome P450 2E1(CYP2E1), both of them can cause DNA damage and cytotoxicity. Recently, the relation between dietary acrylamide and human health has drawn public attention. The purpose of the present study is to investigate the potential mechanism of acrylamide at lower doses on proliferation of human hepotocacinoma Hep G2 cells, and to test whether curcumin has antagonistic effects against acrylamide.Method: MTT assay was used to evaluate the effect of lower doses of acrylamide or combination of curcumin on Hep G2 cells proliferation. The expression of mi R-21 was detected by q RT-PCR. Ed U fluorescence staining was performed to observe the replication of DNA in Hep G2 cells. Flow cytometric assay was applied to investigate the apoptosis of the cells. The expression of proliferation-related proteins was detected by Western Blot.Results:1. The expression of mi R-21 in HepG2 was up-regulated by acrylamide. Lower concentration of acrylamide(≤100μΜ) significantly increased the proliferation of Hep G2 cells and the level of mi R-21. The level of mi R-21 was significantly elevated when administrated 100 μM of acrylamide in Hep G2.2. Acrylamide inhibited PTEN expression and activated the signaling pathway of PTEN/PI3K/AKT. Lower concentration of acrylamide inhibited the expression of PTEN and induced the proliferation-related proteins, such as EGFR, cyclin D1, which contributed to cell proliferation. PI3K/AKT inhibitor, LY294002, decreased the expression of p-AKT, and inhibited the proliferation of Hep G2 cells.3. mi R-21 inhibitor decreased the proliferation of Hep G2 cells. The expression level of mi R-21 in Hep G2 cells was downregulated by mi R-21 inhibitor. The results of colony formation assay and Ed U fluorescence staining indicated that mi R-21 inhibitor could inhibit the proliferation of Hep G2 cells. In addition, CYP2E1 was downregulated by mi R-21 mimic and up-regulated by mi R-21 inhibitor.4. The proliferation of HepG2 cell induced by acrylamide was reduced by curcumin. Flow cytometric assay was used to detect the apoptosis of Hep G2 cells induced by curcumin(10μM). Curcumin effectively reduced acrylamide-induced proliferation, as well as the expression of mi R-21.Conclusion: Acrylamide promoted the expression of mi R-21 in Hep G2 cells. All these results suggest that lower concentration of acrylamide may contribute to progression of hepatocellular carcinoma(HCC). Curcumin could prevent acrylamide triggering this effect.Objective: About 90% of acrylamide and alcohol in the body is metabolized in the liver. Both acrylamide and alcohol possess similar toxic effects on human body. The purpose of the present study was to explore the protective effect of curcumin on BRL cells proliferation induced by the combination of acrylamide and alcohol.Methods: MTT assay was performed to evaluate the pro-proliferation effect of acrylamide and alcohol in combination. Flow cytometry assay analysed the ROS level and cell cycle in cells. The expression of protein was detected by Western Blot. The level of mi R-21 was examined by qRT-PCR.Results: Combination of acrylamide and alcohol at low doses increased cell activity, promoted proliferation and up-regulated the expression of cyclin D1/B1. In addition, the level of mi R-21 was time-dependently increased in BRL cells exposed to low concentration of acrylamide and alcohol. Curcumin could inhibit the combined effects of acrylamide and alcohol on BRL cells.Conclusions: Curcumin suppressed the synergistic effect of acrylamide and alcohol on cell proliferation and cyclin D1/B1 up-regulation in BRL cells.