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| Effects of LAMC1 gene silencing on proliferation and cell cycle of bladder cancer cells |
| LI Binqi1 LI Wenming2▲ |
1.Department of General, Handan Central Hospital, Hebei Province, Handan 056001, China;
2.Pre-hospital Emergency Aid, Handan Central Hospital, Hebei Province, Handan 056001, China |
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Abstract Objective To investigate the expression of recombinant laminin gamma 1 (LAMC1) in bladder cancer cells and the effect of silencing lamc1 gene expression on the proliferation and cell cycle of bladder cancer cells. Methods Cancer and adjacent tissues of 48 patients with bladder cancer admitted to Handan Central Hospital, Hebei Province from January 2019 to December 2021 were selected. RT-qPCR was used to detect the expression of LAMC1 mRNA in UMUC3, T24, 5637, and J82 cells. LAMC1-siRNA plasmid was transfected into 5637 cells, LAMC1 silencing group and negative control group were set up, and the transfection efficiency was verified by RT-qPCR and Western blot. CCK-8 assay, clonogenesis assay and flow cytometry were used to detect the effect of LAMC1 gene silencing on the proliferation, clonogenesis ability, and cell cycle of bladder cancer cells. Western blot was used to detect the expression levels of cyclin A, cyclin D1, p16, and p21. Results The expression level of LAMC1 mRNA in 5637 cells was higher than that in UMUC3, T24, and J82 cells (P＜0.05). There was no significant difference in LAMC1 positive expression rate between bladder and paracancer tissues (P＞0.05). LAMC1 mRNA and protein expression levels in LAMC1 silencing group were lower than those in negative control group (P＜0.01). Cell proliferation and clonogenesis in LAMC1 silencing group were lower than those in negative control group (P＜0.01). The percentage of S phase cells in LAMC1 silencing group was higher than that in negative control group (P＜0.01), and the percentage of G1 phase cells was higher than that in negative control group (P＜0.01). The expression levels of cyclin D1 and cyclin A proteins in LAMC1 silencing group were highly lower than those in negative control group (P＜0.01), while the expression levels of p16 and p21 proteins were higher than those in negative control group (P＜0.01). Conclusion Silencing LAMC1 gene expression can inhibit the expression of cell cycle related proteins, thus inhibiting the proliferation of bladder cancer 5637 cells, suggesting that LAMC1 may be a potential therapeutic target for bladder cancer.
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