Study On The Expression Of SNORD82 In Hepatocellular Carcinoma Cells

The main purpose of this experimental study was to detect the expression of SNORD82 in hepatocellular carcinoma cells,and to test the correlation between the expression level of SNORD82 and the invasive ability of hepatoma cells,and to construct a stable hepatocellular carcinoma cell line with stable expression of SNORD82 using lentivirus transfection technology for subsequent functional studies.The experimental and target gene principle of action provide theoretical basis and reference.Methods:1.q RT-PCR detection of HCCLM3,Hep G2,Huh-7 and HL-7702 SNORD82 expression levels;2.Construction of a lentiviral vector that inhibits the expression of SNORD82,packaging and infecting highly expressed hepatoma cell lines;3.The lentiviral transfection efficiency was observed by fluorescence microscopy,and the expression level of SNORD82 was detected by q RT-PCR.Results:1.q RT-PCR results showed that SNORD82 was detected in both normal hepatocellular carcinoma cell line(HL-7702)and hepatocellular carcinoma cell line(HCCLM3> Hep G2> Huh-7).The relative expression level of SNORD82 in HCCLM3 cell line was 5.580±0.121,that in Hep G2 cell line was 3.521±0.130,that in Huh-7 cell line was 1.417±0.120,and in normal hepatocytes(HL-7702)The relative expression level was 0.924±0.142.There was significant difference between the two groups(P <0.05).The expression levels of SNORD82 in all four cell lines from high to low were as follows: HCCLM3> Hep G2> Huh-7> HL-7702.2.Take SNORD82-RNAi lentivirus with a titer of 4 × 108 TU / ml and then infect liver cancer cell lines and observe its transfection efficiency by fluorescence microscopy to obtain stable hepatoma cell lines.The cell line infected with SNORD82-RNAi lentivirus was defined as KD-HCCLM3 and the negative control virus-infected cell line was defined as NC-HCCLM3.The data of SNORD82 in three hepatoma cells(HCCLM3,NC-HCCLM3,KD-HCCLM3)detected by real-time fluorescence quantitative PCR showed that the relative expression level of SNORD82 in HCCLM3 cells was(2.51 ± 0.18),and the relative expression level in NC-2.38 ± 0.21).The relative expression level in KD-HCCLM3 group was(0.527 ± 0.36).There was no significant difference between HCCLM3 group and NC-HCCLM3 group(P> 0.05).Compared with HCCLM3 and NC-HCCLM3 groups,the expression of KD-HCCLM3 group was significantly decreased(P <0.05).Conclusion:1.SNORD82 is upregulated in hepatoma cells and positively correlated with cell invasion and metastasis.2.Lentivirus transfection was used to inhibit the expression of SNORD82 in HCCLM3 and a lentivirus cell line was successfully constructed.