Effect Of LncRNA LOC730101 On Proliferation, Migration And Invasion Of Hepatocellular Carcinoma

Objective:In this paper,we explored the expression of long non-coding RNA（lncRNA）LOC730101 in hepatocellular carcinoma（HCC）tissues and corresponding adjacent tissues,hepatocellular carcinoma cell lines and normal liver cell lines,and studied the effect of lncRNA LOC730101 on the proliferation,migration and invasion of hepatocellular carcinoma,hoping to find new hepatocellular carcinoma markers and therapeutic targets.Methods:1.Firstly,the dysregulated lncRNAs in hepatocellular carcinoma were retrieved from TCGA,LncCAR,NCBI and other databases.Combined with bioinformatics analysis and literature reading,the research object—lncRNA LOC730101 was determined.The expression of lncRNA LOC730101 in 374liver cancer tissues and 50 normal tissues in LIHC data set of TCGA database was analyzed by R language software.2.The cancer tissues and corresponding adjacent tissues of 40 patients with HCC diagnosed by pathology for the first time in the Affiliated Tumor Hospital of Guangxi Medical University in 2018（only undergoing surgery,without radiotherapy and chemotherapy）were collected.The expression level of LOC730101 in the samples was detected by q RT-PCR,and the expression difference was analyzed by paired sample t-test.The relative expression levels of LOC730101 in hepatocellular carcinoma cell lines Huh-7,HCCLM3,Hep G2 and human normal liver cell line L02 were detected by q RT-PCR,and independent sample t-test was used for statistical analysis.3.The stable cell lines overexpressing LOC730101（OE-LOC730101）and knocking down LOC730101（sh-LOC730101-LOC730101 RNAi 105959,LOC730101 RNAi 105960,LOC730101 RNAi 105961）were constructed by lentivirus transfection in hepatocellular carcinoma cell lines Huh-7 and HCCLM3.The stably transfected cells were screened by puromycin,and the transfection efficiency was verified by q RT-PCR.The target with the most significant knockdown effect of LOC730101 was selected.Finally,blank control group and negative control group（sh-NC,OE-NC）were set up for each cell line.CCK-8 assay was used to detect the proliferation ability of cells in each group.Scratch migration assay and transwell invasion assay were used to detect the migration and invasion ability.Results:1.The results of TCGA database analysis showed that the expression level of LOC730101 in HCC tissues was higher than that in normal tissues（P<0.001）.The results of q RT-PCR showed that the expression of LOC730101 in 40cases of HCC tissues was significantly higher than that in adjacent tissues（P<0.001）.The expression level of LOC730101 in liver cancer cell lines（Huh-7,Hep G2,HCCLM3）was higher than that in human normal liver cell line L02（all P<0.05）,indicating that LOC730101 has research value in hepatocellular carcinoma.2.The results of q RT-PCR showed that there was no significant difference in the expression level of LOC730101 between the blank control group（control group）and the negative control group（sh-NC,OE-NC）.Compared with the sh-NC group,the expression level of LOC730101 in LOC730101 RNAi（105960）was significantly decreased（P<0.01）.Compared with the OE-NC group,the expression level of LOC730101 in the OE-LOC730101 group was significantly increased（P<0.01）.The above results showed that the stable cell lines with overexpression and knockdown of LOC730101 were successfully constructed.3.CCK-8 results of two hepatoma cells:In Huh-7,the proliferation ability of the sh-LOC730101 group was inhibited(0.981±0.021 vs 0.825±0.034,P_72h=0.002)compared with the sh-NC group,and the OE-LOC730101 group was increased(1.097±0.004 vs 1.471±0.096,P_72h<0.001)compared with the OE-NC group.In HCCLM3,the proliferation ability of the sh-LOC730101group was significantly decreased(0.885±0.037 vs 0.571±0.108,P_72h<0.001)compared with the sh-NC group,and the OE-LOC730101 group was increased(0.972±0.105 vs 1.233±0.046,P_72h=0.004)compared with the OE-NC group.4.Scratch migration assay results of two hepatoma cells:In Huh-7,the migration ability of the sh-LOC730101 group was significantly lower than that of the sh-NC group（0.433±0.054 vs 0.265±0.026,P=0.002）,and the OE-LOC730101 group was significantly enhanced（0.381±0.054 vs 0.552±0.049,P=0.002）compared with the OE-NC group.In HCCLM3,the migration ability of the sh-LOC730101 group was decreased（0.275±0.036 vs0.197±0.015,P=0.011）compared with the sh-NC group,and the OE-LOC730101 group was significantly higher（0.287±0.029 vs 0.432±0.040,P<0.001）than that of the OE-NC group.5.Transwell invasion assay results of two hepatoma cells:In Huh-7,the invasion ability of the sh-LOC730101 group decreased（212±20 vs 122±16,P=0.002）compared with the sh-NC group,and the OE-LOC730101 group was significantly increased（169±17 vs 285±22,P<0.001）compared with the OE-NC group.In HCCLM3,the invasion ability of the sh-LOC730101 group was significantly lower than that of the sh-NC group（214±20 vs 82±9,P<0.001）,and the OE-LOC730101 group was significantly enhanced（224±24vs 333±35,P<0.001）compared with the OE-NC group.Conclusion（s）:1.We found that lncRNA LOC730101 was highly expressed in hepatocellular carcinoma tissues and cell lines.2.The up-regulated expression of LOC730101 in HCC cell may promote its proliferation,migration and invasion.3.Lnc RNA LOC730101 is expected to become a new tumor marker and therapeutic target for hepatocellular carcinoma,and the mechanism in hepatocellular carcinoma needs further study.