Differential Expression Of CircRNA In Intrahepatic Cholangiocarcinoma And The Effect Of Circ＿0089153 On Proliferation,migration And Invasion Of ICC Cells

Background & objective: The abnormal expression of circular RNA(circRNA)is closely related to the occurrence and development of malignant tumors.The purpose of this study was to investigate the abnormal expression of circRNA in intrahepatic cholangiocarcinoma(ICC)and the effect of circ＿0089153 on the proliferation,migration and invasion of ICC tumor cel s.Methods: The specimens of 5 patients with ICC confirmed by pathology in Fujian Provincial Hospital were used to screen the differentially expressed circRNA,in ICC cancer tissues and corresponding paracancerous tissues(FC > 2 times,t-test P< 0.05)by high-throughput circRNA microarray technique.The circ＿0089153 with the most significant multiple changes in length 500-1500 bp,FC > 4 times and P <0.001,raw intensity > 300 were selected for real-time quantitative PCR verification.The expression of circ＿0089153 in HCCC-9810 and RBE cell lines was overexpressed by lentivirus.The effects of circ＿0089153 on the proliferation,migration and invasion of ICC cells were detected by CCK8 test,streak test,Transwell test and flow apoptosis assay.Results: Compared with the normal tissues adjacent to cancer,there were seventy-six differentially expressed circRNA in ICC tissues,of which sixty-three were higher and thirteen were lower in ICC than those in paracancerous tissues.Further improve the conditions(a.FC > 4 times and P < 0.001,b.the raw intensity of each group is more than 300;c.the length of each circRNA is between500-1500bp),ten circRNAs were screened out,of which eight were up-regulated(circ＿0001402,circ＿0002886,circ＿0007750,circ＿0006827,circ＿0044226,circ＿0001756,circ＿0001359,circ＿0062682),two down-regulated(circ＿0048764,circ＿0089153),especially circ＿0089153 multiple change(FC=103.2137475).Its relative expression in ICC tissues(7.8920 ±1.0673)was significantly lower than that in normal tissues adjacent to cancer(14.5815 ±0.7369)(P < 0.0001).Real-time quantitative PCR was further verified,which was consistent with the results of gene chip.The results of CCK8 experiment showed that the proliferative activity of ICC cells in circ＿0089153 overexpressed group(HCCC9810: 114.8159 ±5.1381;RBE:117.2350 ±7.0792)was significantly higher than that in unloaded group(HCCC9810: 100.0000 ±4.8970;RBE:100.0000 ±14.8965).The results of flow cytometry apoptosis assay showed the apoptosis rate of ICC cells in circ＿0089153overexpressed group(HCCC9810: 11.2767 ±0.2434;RBE:4.5267 ±0.3009)and empty group(HCCC9810:16.6733 ±0.6603).Compared with RBE:6.7933±0.5341),it decreased significantly(P < 0.05).The results of cell marking experiment showed that the migration ability of ICC cells in circ＿0089153overexpressed group(HCCC9810: 9.0702 ±1.0902;RBE:6.1970 ±0.6873)was significantly higher than that in unloaded group(HCCC9810: 6.2065 ±1.1974;RBE:4.1978 ±1.1701).The results of cell invasion assay showed the invasive ability of ICC cells in circ＿0089153 overexpressed group(HCCC9810: 0.3696±0.0258;RBE:0.2967 ±0.0157)and empty group(HCCC9810:0.2743±0.0142).Compared with RBE:0.2498 ±0.0188),it was significantly increased(P< 0.05).Conclusion: There are many kinds of abnormal expression of circRNA,circ＿0089153 in ICC tissues and low expression in ICC tissues.Overexpression of circ＿0089153 gene can promote the growth,migration,invasion and inhibit apoptosis of ICC.