Construction of lentiviral vector carrying miR -122 and function of miR -122 in hepatocellular carcinoma cells.

Objective To construct the lentiviral vector carrying miR - 122 ( Lv - miRl 22 ) and investigate the function of stably expressed miR - 122 in hepatocellular carcinoma ( HCC) cells. Methods Pre - miR - 122 was cloned into lentiviral vector pLUNIG to obtain the Lv - miR122 that had stable expression of miR - 122 ; Lv - miR122 was packaged in 293T cells along with other three vectors pRRE, pRSV -REV, and pCMV - VSVG, and virus supernatant was obtained by concentration; the virus supernatant was used to infect HCC cell line HepG2. The expression of miR - 122 was measured by Q - RT - PCR. The proliferation of HepG2 cells was analyzed by MTT assay and col-ony formation assay. Blank lentiviral vector ( Lv - Ctrl) - infected HepG2 cells and uninfected HepG2 cells were used as controls. Results The expression of miR - 122 was significantly higher in Lv - miR122 - infected HepG2 cells than in Lv - Ctrl - infected HepG2 cells ( t -- 10. 745, P < 0. 01 ). The Lv - miR122 - infected HepG2 cells ( with stable expression of miR - 122 ) had significantly suppressed prolifera-tion compared with Lv - Ctrl - infected and uninfected HepG2 cells (P <0. 05 for both comparisons) ; moreover, miR - 122 significantly in-hibited the colony formation of HepG2 cells ( t -- 5. 256, P < 0. 01 ) . Conclusion The Lv - miR122 can be successfully constructed, and the carried miR -122 can be stably expressed and can inhibit the proliferation of HepG2 cells. [ABSTRACT FROM AUTHOR]