Effects Of LncRNA NBAT1 On The Biological Behavior Of Bladder Cancer 5637 Cells And The Construction Of Its Lentiviral Vector

Objective: To investigate the expression of long non-coding RNA NBAT1(neuroblastoma associated transcript 1)in human bladder cancer cells,and to explore the effects of downregulation of NBAT1 gene expression on the biological behavior of bladder cancer 5637 cells.By constructing shRNA lentiviral vector,the specific function and related mechanism of NBAT1 gene in bladder cancer cells will be further studied.Methods: The mRNA expression level of NBAT1 in seven cell lines of human bladder cancer(RT,5637,EJ,J82,TCC-SUP,T24,and UM-UC-3)was detected by real-time fluorescent quantitative PCR(RT-qPCR),and the correlation of NBAT1 expression with the malignancy of bladder cancer cells was analyzed.The specific siRNA targeting NBAT1 gene was transfected into bladder cancer 5637 cells,and the change of NBAT1 gene expression was detected by RT-qPCR.Then the effects of NBAT1 gene silencing on the viability,migration,invasion and cisplatin-induced apoptosis of 5637 cells were analyzed by using CCK-8 assay,wound healing assay,Transwell assay and Hoechst assay,respectively.The NBAT1 gene and the negative control lentiviral sh RNA vector were constructed and packaged into virus particles,by which the target cells were infected.Results: The expression level of NBAT1 mRNA in bladder cancer RT4 cells with lower invasive potential was significantly higher than that in bladder cancer cells(5637,EJ,J82,TCC-SUP,T24,and UM-UC-3)with higher invasive potential(all P < 0.01).After the specific siRNA targeting NBAT1 gene was transfected into bladder cancer 5637 cells,the expression of NBAT1 gene was significantly inhibited(P < 0.01).Furthermore,the viability,migration and invasion of 5637 cells were significantly promoted(all P < 0.01),but cisplatin-induced apoptosis was not significantly changed after NBAT1 gene silencing(P > 0.05).The constructed vector was identified by colony PCR assay and sanger sequencing assay,by which the DNA sequence was confirmed with no mutations.The constructed vector was packaged into a virus to infect the target cells.Conclusion: The expression level of NBAT1 gene may be correlated with the malignant degree of bladder cancer cells.Downregulation of NBAT1 gene expression would increase the cell viability,and promote the migration and invasion of bladder cancer 5637 cells.The lentiviral shRNA vector was constructed successfully and had the ability to infect the target cells.