claudin-1下調對膽囊癌SGC996細胞生理過程的影響

2021年8月，蚌埠醫學院**附屬醫院肝膽外科；東南大學中大醫院普外科(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老師研究團隊在《Oncology Letters》上發表論文：

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

“claudin-1下調對膽囊癌SGC996細胞生理過程的影響”

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：

膽囊癌復發率高，死亡率高，治療選擇有限。因此，闡明這種疾病的潛在分子機制將有助于實現早期診斷和潛在地確定新的治療靶點。Claudin-1是一種緊密連接蛋白，與多種癌癥的發生和預后相關，我們的初步研究表明，Claudin-1在膽囊癌組織中的表達升高。因此，本研究旨在探討下調claudin-1對膽囊癌細胞生理過程的影響。以claudin-1- rna干擾慢病毒(LV-CLDN1-RNAi)轉染膽囊癌SGC996細胞株，下調claudin-1的表達，研究其對細胞增殖、細胞周期、細胞凋亡和細胞侵襲的下游影響。轉染LV-CLDN1-RNAi后，MTT檢測結果顯示下調claudin-1不影響SGC996細胞的增殖。然而，流式細胞術分析顯示，G1期的細胞數量明顯增加，而S期的細胞數量明顯減少。Annexin V-APC單色染色顯示，下調cladin -1表達增加了細胞凋亡比例，western blot分析結果證實了這一點，其中促凋亡的b細胞淋巴瘤2 (Bcl-2)相關X蛋白和抗凋亡的Bcl-2蛋白水平分別升高和降低。最后，Transwell實驗表明，claudin-1下調抑制細胞侵襲。綜上所述，本研究結果提示下調claudin-1表達可促進膽囊癌細胞凋亡，抑制細胞侵襲，提示claudin-1可能參與膽囊癌的復發轉移。這些發現為考慮claudin-1作為膽囊癌治療的新靶點提供了理論和實驗基礎。

該論文中，膽囊癌SGC996和GBC-SD細胞系以及膽管癌QBC939細胞的體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.