Effects Of Long Non-coding RNA H19 Expression On Cell Proliferation,Migration,Invasive Ability And Drug Resistance Of Cholangiocarcinoma Cells

Background and objective:Cholangiocarcinoma,a soild malignant tumor that originating from bile duct epithelial cells,is accounting for about 2% to 3% of the digestive tract tumors.Although the incidence of cholangiocarcinoma is low,but its global incidence has increased year by year.In recent years,surgical techniques,surgical methods and perioperative management have been greatly developed and improved,but the relevant mortality rate of cholangiocarcinoma is still increased by 39%.The cause seems to related with the concealed pathogenesis,special anatomical site,diversified way of transfer and chemotherapy-resistant.Recent studies have shown that long non-coding RNA H19,one of the earliest genes that has been shown to have imprinting properties,plays an important role in normal embryonic development and tumors occurrence and progression.H19 not only benefit for carcinomatosis,but also has the effect to cancer inhibition.H19 was overexpression in gastric cancer,esophageal squamous cell carcinoma,bladder cancer and osteosarcoma,while low expression in hepatocellular carcinoma,nephroblastoma and prostate cancer.So far,the role of H19 in cholangiocarcinoma is rarely reported.In this study,we investigated the effect o f high expression of H19 on the proliferation,migration,invasion and drug resistance of cholangiocarcinoma cell line QBC939 by constructing high expression of H19 and infecting QBC939 cells.Methods:1.The expression of H19 in 18 cases of cholangiocarcinoma and adjacent normal bile duct was detected by RT-PCR.2.Construction of H19 high expression of chronic virus and infection of QBC939 cells,and then use CCK-8 kit,scratch test and Transwell chamber to detected the relationship between H19 high expression and cell proliferation,migration and invasion.3.To investigate the sensitivity of QBC939,which overexpression H19,to gemcitabine in chemotherapy chemotherapy by CCK-8 drug toxicity test and nude mouse tumorigenesis test.Results:1.In this study,the expression of H19 in 18 cases of cholangiocarcinoma and normal paracancerous bile duct was studied,and detected that the expression of H19 in cholangiocarcinoma was significantly lower than that in adjacent normal bile duct tissues(4.03±1.02 VS 2.99±1.11,p <0.05);2.CCK-8 method showed that there was no significant difference between the high expression group and the empty plasmid group in cell proliferation rate at 48 h(0.40 ± 0.05 vs 0.31 ± 0.08,p> 0.05),but the cell proliferation rate of H19 high expression group was significantly lower than that of empty plasmid cells at 72 h and 96h(0.63 ± 0.05 vs 0.40 ± 0.06 and 0.74 ± 0.05 vs 0.54 ± 0.04),the difference was statistically significant(p <0.05)3.The results of scratches test showed that the scribing width of H19 high expression group and empty plasmid control group at 0 hour was 808 ± 56 um and 816 ± 48 um,respectively.There was no significant difference between the two groups(p> 0.05)At 24 hour and 48 hour,the width of the two groups was 632 ± 67 um VS 400 ± 56 um,482 ± 40 um VS 213 ± 66 um.The difference between the two groups were statistically significant(p <0.05)4.Transwell assay showed that the number of perforated cells in H19 overexpression group was significantly lower than that in empty plasmid group(66.80 ± 11.99 vs 173.6 ± 37.77),and there was significant difference between the two groups(p <0.05).5.CCK-8 method showed that there was no significant difference in the survival rate between H19 high expression group and empty plasmid group(0.5.16 ± 4.43 vs 79.91 ± 6.59;75.76 ± 8.48,p> 0.05)at the gemcitabine concentration of 0.5ug/ml and 1.0ug /ml;However,undergo the concentration of 2ug/ml,5ug/ml and 10ug/ml,the survival rate of H19 high expression group was obviously lower than the empty plasmid group(67.04±5.33 VS 56.28±8.25;54.21±3.66 VS 33.51±6.29,47.08±2.20 VS 25.31±8.53),and here was significant difference between the two groups(p <0.05).6.The results of transplanted tumor showed that the weight of xenograft in H19 high expression group was significantly lower than that in empty plasmid group(0.72 ± 0.05 vs 0.60 ± 0.03 0.53 ± 0.05 vs 0.35 ± 0.05),and the difference between the two groups was statistically significant(p < 0.05).Conclusions:The results showed that long non-coding RNA H19 was low expression in cholangiocarcinoma,and the up-regulation of H19 expression reduced the proliferation,migration and invasion of cholangiocarcinoma cells and the resistance to gemcitabine.It was confirmed that H19 could act as a tumor suppressor gene in cholangiocarcinoma.