claudin-1下調(diào)對(duì)膽囊癌SGC996細(xì)胞生理過(guò)程的影響

2021年8月，蚌埠醫(yī)學(xué)院**附屬醫(yī)院肝膽外科；東南大學(xué)中大醫(yī)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(tuán)隊(duì)在《Oncology Letters》上發(fā)表論文：

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

“claudin-1下調(diào)對(duì)膽囊癌SGC996細(xì)胞生理過(guò)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(fù)發(fā)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(jī)制將有助于實(shí)現(xiàn)早期診斷和潛在地確定新的治療靶點(diǎn)。Claudin-1是一種緊密連接蛋白，與多種癌癥的發(fā)生和預(yù)后相關(guān)，我們的初步研究表明，Claudin-1在膽囊癌組織中的表達(dá)升高。因此，本研究旨在探討下調(diào)claudin-1對(duì)膽囊癌細(xì)胞生理過(guò)程的影響。以claudin-1- rna干擾慢病毒(LV-CLDN1-RNAi)轉(zhuǎn)染膽囊癌SGC996細(xì)胞株，下調(diào)claudin-1的表達(dá)，研究其對(duì)細(xì)胞增殖、細(xì)胞周期、細(xì)胞凋亡和細(xì)胞侵襲的下游影響。轉(zhuǎn)染LV-CLDN1-RNAi后，MTT檢測(cè)結(jié)果顯示下調(diào)claudin-1不影響SGC996細(xì)胞的增殖。然而，流式細(xì)胞術(shù)分析顯示，G1期的細(xì)胞數(shù)量明顯增加，而S期的細(xì)胞數(shù)量明顯減少。Annexin V-APC單色染色顯示，下調(diào)cladin -1表達(dá)增加了細(xì)胞凋亡比例，western blot分析結(jié)果證實(shí)了這一點(diǎn)，其中促凋亡的b細(xì)胞淋巴瘤2 (Bcl-2)相關(guān)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(shí)驗(yàn)表明，claudin-1下調(diào)抑制細(xì)胞侵襲。綜上所述，本研究結(jié)果提示下調(diào)claudin-1表達(dá)可促進(jìn)膽囊癌細(xì)胞凋亡，抑制細(xì)胞侵襲，提示claudin-1可能參與膽囊癌的復(fù)發(fā)轉(zhuǎn)移。這些發(fā)現(xiàn)為考慮claudin-1作為膽囊癌治療的新靶點(diǎn)提供了理論和實(shí)驗(yàn)基礎(chǔ)。

該論文中，膽囊癌SGC996和GBC-SD細(xì)胞系以及膽管癌QBC939細(xì)胞的體外培養(yǎng)是使用Ausbian特級(jí)胎牛血清完成的。欲了解或購(gòu)買(mǎi)Ausbian特級(jí)胎牛血清可以聯(lián)系北京締一生物400-166-8600.