Effects of claudin-1 downregulation on the physiological processes of gallbladder cancer SGC996 cells

Gallbladder cancer has a high recurrence and mortality rate, with limited treatment options. Therefore, elucidating the underlying molecular mechanisms of this disease would be beneficial to achieve an earlier diagnosis and potentially identify novel treatment targets. Claudin-1 is a tight junction protein associated with the development and prognosis of several types of cancer, and our preliminary studies have demonstrated that claudin-1 expression is elevated in gallbladder cancer tissues. Therefore, the aim of the present study was to investigate the effects of downregulating claudin-1 on the physiological processes of gallbladder cancer cells. The gallbladder cancer SGC996 cell line was transfected with claudin-1-RNA interference lentivirus (LV-CLDN1-RNAi) to downregulate claudin-1 expression, and the downstream effects on cell proliferation, the cell cycle, apoptosis and cell invasion were investigated. Following transfection with LV-CLDN1-RNAi, the results of an MTT assay revealed that downregulating claudin-1 did not affect the proliferation of the SGC996 cells. However, flow cytometry analysis demonstrated that the number of cells arrested in the G(1) phase increased significantly, whereas the amount of cells arrested in the S phase was significantly reduced. Annexin V-APC single-color staining demonstrated that downregulating claudin-1 expression increased the ratio of cell apoptosis, which was confirmed by the results of western blot analysis, in which levels of the pro-apoptotic B-cell lymphoma 2 (Bcl-2)-associated X protein and anti-apoptotic Bcl-2 protein were increased and decreased, respectively. Finally, a Transwell assay indicated that claudin-1 downregulation inhibited cell invasion. Overall, the results from the present study indicated that downregulating claudin-1 expression promoted the apoptosis of gallbladder cancer cells and inhibited cell invasion, indicating that claudin-1 may be involved in the recurrence and metastasis of gallbladder cancer. These insights provide theoretical and experimental foundations for considering claudin-1 as a novel target for the treatment of gallbladder cancer.