The Effects Of Linc00152 On Proliferation, Invasion And Migration Of Pancreatic Cancer MIA PaCa-2 Cell

Objective: To investigate the effect of long non-coding RNA Linc00152 on the proliferation,invasion and migration of human pancreatic cancer cell line MIA PaCa-2.Methods: Twenty pairs of pancreatic cancer tissues and their corresponding normal adjacent pancreatic tissues were collected from the Department of Cholangiopancreatology in hospital.Human normal pancreatic cell HPDE and pancreatic cancer cell lines MIA PaCa-2,BxPC-3 and Capan2 were purchased from American ATCC cell bank.In our study,we determined the expression levels of Linc00152 in Pancreatic cancer tissues and cell lines(MIA PaCa-2,BxPC-3 and Capan2)by quantitative real time polymerase chain reaction(qRT‐PCR).Human pancreatic cancer cell line MIA PaCa-2 cells were randomly divided into a negative control group(si-NC)and a Linc00152 down-regulated group(si-Linc00152).The siRNA(small interfering RNA)interference model of Linc00152 was designed,and the negative control sequence and siRNA inhibition sequence were transfected into the si-NC group and the si-Linc00152 group,respectively.And the transfection efficiency level of Linc00152 in MIA PaCa-2 cells was measured by RT-qPCR.Firstly,the proliferation of MIA PaCa-2 cells was detected by CCK-8 assay and plate cloning assay.The cell cycle of MIA PaCa-2 cells was detected by flow cytometry.The pancreatic cancer cells MIA PaCa-2 were detected by Western-blot.Cell cycle-associated protein cyclin D1,CDK4 expression: Secondly,Transwell assay and wound healing assay were used to measure the invasion and migration ability of MIA PaCa-2 cells;finally,Western-blot detected MIA PaCa-2 Expression levels of cell-associated proteins vimentin and N-cadherin.Results: Compared with the adjacent tissues,the expression of Linc00152 in pancreatic cancer tissues increased significantly(P<0.001).The expressions of Linc00152 in cell lines(MIA PaCa-2,BxPC-3 and Capan2)of pancreatic cancer were higher than that in humans normal pancreatic cells HPDE,and the highest expression in MIA PaCa-2(P<0.01).The siRNA interference model was successfully designed.RT-q PCR results showed that the expression of Linc00152 in si-Linc00152 group was significantly lower than that in control group after 48 hours of transfection.The cell viability of MIA PaCa-2 was measured at 6 h,24 h,48 h,72 h and 96 h with CCK-8 assay.Compared with the si-NC group,the cells in the si-Linc00152 group showed a significant decrease in proliferation at 48 h,72 h,96 h(P< 0.01).After 8 days of plate cloning,the number of cell clones in the si-NC group was(412±12),while the number of cell clones in the si-Linc00152 group was(226±8).The results showed that the number of cell clones in the si-NC group was significantly higher than that in the si-Linc00152 group(P<0.01).The results of flow cytometry showed that,compared with the si-NC group,the percentage of G0/G1 phase in the cell cycle of MIA PaCa-2 in the si-Linc00152 group increased,which suggesting that cell cycle arrest occurred(P<0.01).Transwell assay showed that the number of cell invasion after silencing Linc00152 was significantly reduced(P<0.01).Transwell assay showed that the number of cell migration after silencing Linc00152 was significantly reduced(P<0.01).Wound healing assay showed that compared with the si-NC group,the invasive ability of the si-Linc00152 group were significantly decreased(P<0.01).Western-blot results showed that the expression levels of cyclin D1 and CDK4 in si-Linc00152 group were significantly lower than those in the control group(P<0.01).Compared with the control group si-NC,the expression levels of vimentin and N-cadherin protein in the si-Linc00152 group were significantly lower(P<0.01),that inhibits Epithelial-Mesenchymal-Transition(EMT)in pancreatic cancer cells MIA PaCa-2.Conclusion: 1.Linc00152 is highly expressed in pancreatic cancer tissues and cell lines.2.Silencing Linc00152 can inhibit the proliferation,invasion and migration in the pancreatic cancer cells MIA PaCa-2.3.Silencing Linc00152 can block the cell cycle of pancreatic cancer cells MIA PaCa-2 at G0/G1 phase,that may be related to cyclinD1-CDK4 pathway.4.Silencing Linc00152 can inhibit EMT of MIA PaCa-2 cells,which may be related to the decreased expression of vimentin and N-cadherin protein.