claudin-1下调对胆囊癌SGC996细胞生理过程的影响

2021年8月，蚌埠医学院**附属医院肝胆外科；东南大学中大医院普外科(Department of Hepatobiliary Surgery;Clinical Laboratory, The First Affiliated Hospital, Bengbu Medical College, Bengbu, Anhui 233004;Department of General Surgery, Zhongda Hospital,Southeast University, Nanjing, Jiangsu 210009, P.R. China) HAO JIN老师研究团队在《Oncology Letters》上发表论文：

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

“claudin-1下调对胆囊癌SGC996细胞生理过程的影响”

Abstract：

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 G1 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.

摘要：

胆囊癌复发率高，死亡率高，治疗选择有限。因此，阐明这种疾病的潜在分子机制将有助于实现早期诊断和潜在地确定新的治疗靶点。Claudin-1是一种紧密连接蛋白，与多种癌症的发生和预后相关，我们的初步研究表明，Claudin-1在胆囊癌组织中的表达升高。因此，本研究旨在探讨下调claudin-1对胆囊癌细胞生理过程的影响。以claudin-1- rna干扰慢病毒(LV-CLDN1-RNAi)转染胆囊癌SGC996细胞株，下调claudin-1的表达，研究其对细胞增殖、细胞周期、细胞凋亡和细胞侵袭的下游影响。转染LV-CLDN1-RNAi后，MTT检测结果显示下调claudin-1不影响SGC996细胞的增殖。然而，流式细胞术分析显示，G1期的细胞数量明显增加，而S期的细胞数量明显减少。Annexin V-APC单色染色显示，下调cladin -1表达增加了细胞凋亡比例，western blot分析结果证实了这一点，其中促凋亡的b细胞淋巴瘤2 (Bcl-2)相关X蛋白和抗凋亡的Bcl-2蛋白水平分别升高和降低。最后，Transwell实验表明，claudin-1下调抑制细胞侵袭。综上所述，本研究结果提示下调claudin-1表达可促进胆囊癌细胞凋亡，抑制细胞侵袭，提示claudin-1可能参与胆囊癌的复发转移。这些发现为考虑claudin-1作为胆囊癌治疗的新靶点提供了理论和实验基础。

该论文中，胆囊癌SGC996和GBC-SD细胞系以及胆管癌QBC939细胞的体外培养是使用Ausbian特级胎牛血清完成的。欲了解或购买Ausbian特级胎牛血清可以联系北京缔一生物400-166-8600.