Effect of LINC00665 on hepatocellular carcinoma cell proliferation, apoptosis, migration and invasion by targeting miR-379-5p / 中华肝胆外科杂志

ABSTRACT

Objective:To investigate the effect and mechanism of LINC00665 on hepatocellular carcinoma cell proliferation, migration, invasion and apoptosis.Methods:From June 2013 to June 2018, 126 liver cancer tissue and adjacent tissue (more than 2 cm from the edge of liver cancer tissue) specimens were collected in the Third People's Hospital of Jinan, 86 male and 40 female were included, aged 25.0-72.0 (48.2±9.9) years. The expression level of LINC00665 in 126 liver cancer tissues and adjacent tissues were detected by qRT-PCR. The survival rate of hepatocellular carcinoma HCC9204 cells was determined by CCK8 assay. The apoptosis of HCC9204 cells was detected by flow cytometry, cell migration and invasion were detected by Transwell assay. And the dual-luciferase reporter assay system was implemented to investigate the correlations between LINC00665 and miR-379-5p.Results:Compared with the adjacent tissues group, the expression level of LINC00665 in liver cancer tissues group was increased (1.00±0.10 vs. 1.82±0.18), with statistically significant difference ( P<0.05). When LINC00665 was inhibited [(1.01±0.10) vs. (0.53±0.05)], the expression of Cyclin D1, B-cell lymphoma-2(Bcl-2) and matrix metalloproteinase-2(MMP-2) in HCC9204 cells were decreased as well [(0.74±0.07)vs. (0.13±0.01); (0.81±0.08) vs. (0.35±0.03); (0.69±0.07) vs. (0.22±0.02)], but the level of p21, Bax and E-cadherin were increased [(0.20±0.02) vs. (0.66±0.06); (0.26±0.03) vs. (0.78±0.08); (0.17±0.02) vs. (0.72±0.07)], cell survival rate was decreased [(100.13±10.14)% vs. (46.22±4.73)%], apoptosis rate was increased [(8.42±0.91)% vs. (23.34±2.37)%], and the number of migrating and invading cells were decreased [(109±11) vs. (53±5); (101±10) vs. (47±5)], the differences were statistically significant (all P<0.05). In miR-379-5p overexpression group, the relative activity of firefly luciferase in cells which were co-transfected with wild-type WT-LINC00665 report vector was significantly decreased [(1.03±0.10) ratio (0.24±0.02)], and the relative activity of luciferase in cells which were transfected with mutant MUT-LINC00665 was not decreased significantly ( P>0.05); The level of miR-379-5p in the LINC00665 overexpression group (pcDNA3.1-LINC00665) was decreased [(1.01±0.10) vs (0.37±0.04)]; but was increased in the LINC00665 inhibition group (si-LINC00665)[(0.98±0.10) vs (1.66±0.17)], with statistically significant differences ( P<0.05); Decreasing the content of LINC00665 and miR-379-5p, The cell survival rate was increased [(46.53±4.72)% vs. (82.26±8.34)%], the apoptosis rate was decreased (23.51±2.44)% vs. (12.07±1.21)%], and the number of migrating cells and invasive cells were increased [(54±6) vs. (92±9); (48±5) vs. (88±9)] when LINC00665 and miR-379-5p were inhibited the difference were statistically significant (all P<0.05). Conclusions:In hepatocellular carcinoma HCC9204, LINC00665 targets the regulation of miR-379-5p expression, thereby regulating the proliferation, migration, invasion, and apoptosis of hepatocellular carcinoma HCC9204, which is a potential molecular target for liver cancer.