Department of General Surgery, Nanjing Medical University, Affiliated Suzhou Hospital, Suzhou Municipal Hospital, Gusu, Suzhou, Jiangsu 215000, P.R. China.
J BUON. 2020 Mar-Apr;25(2):1251-1256.
The main focus of the current research work was to unveil the anticancer activity of the naturally occurring Sinensetin flavone against aggressive gall bladder cancer adenocarcinoma (GBAC) TJ-GBC2 cell line. Its effect of inducing apoptosis mediated via targeting PTEN/PI3K/AKT signalling pathway were also examined along with cell migration and invasion.
Cell proliferation was tested by MTT cell viability assay. Fluorescence microscopy was utilized to carry out apoptosis related studies via DAPI staining along with flow cytometry using annexin V/propidium iodide (PI) assay. Further, western blotting analysis was carried out to examine the effects of Sinensetin on the expressions of apoptosis-related proteins and Bax Bcl-2 along with PTEN/PI3K/AKT signalling pathway. The impact of the test molecule on cell migration and invasion was studied through wound healing assay and transwell cell invasion assay respectively.
The results showed that Sinensetin treatment caused a significant retardation in cell viability, in a dose-dependent fashion. DAPI staining assay and annexin V/PI assay revealed that the cell viability of GBC cells was retarded due to induction of apoptosis. It was also associated with downregulation of Bcl-2 and upregulation of Bax levels. Further, wound healing assay and transwell cell invasion assay revealed that cell migration as well as cell invasion of cancer gallbladder cells was decreased in a concentration-dependent fashion. It was further seen that Sinensetin treatment resulted in inhibition of matrix metalloproteinase (MMP)-2 and enhancement of MMP-9 protein expressions. Results also showed that the tested molecule had the potential to inhibit PTEN/PI3K/AKT signalling pathway.
In conclusion, the current study indicated that Sinensetin flavone has the potential to be developed as a candidate drug against gallbladder adenocarcinoma provided more toxicological and in vivo studies are carried out.
本研究工作的主要重点是揭示天然存在的橙皮素黄酮对侵袭性胆囊腺癌（GBAC）TJ-GBC2 细胞系的抗癌活性。还研究了其通过靶向 PTEN/PI3K/AKT 信号通路诱导细胞凋亡的作用以及细胞迁移和侵袭。
通过 MTT 细胞活力测定法测试细胞增殖。荧光显微镜用于通过 DAPI 染色进行与凋亡相关的研究，并通过使用 Annexin V/碘化丙啶（PI）测定法的流式细胞术进行研究。此外，通过 Western blot 分析研究橙皮素对凋亡相关蛋白和 Bax/Bcl-2 的表达以及 PTEN/PI3K/AKT 信号通路的影响。通过划痕愈合试验和 Transwell 细胞侵袭试验分别研究测试分子对细胞迁移和侵袭的影响。
结果表明，橙皮素处理以剂量依赖性方式导致细胞活力显著延迟。DAPI 染色试验和 Annexin V/PI 测定法显示，GBC 细胞的细胞活力因诱导细胞凋亡而延迟。这也与 Bcl-2 下调和 Bax 水平上调有关。此外，划痕愈合试验和 Transwell 细胞侵袭试验表明，癌症胆囊细胞的细胞迁移和细胞侵袭以浓度依赖性方式减少。进一步观察到，橙皮素处理导致基质金属蛋白酶（MMP）-2 的抑制和 MMP-9 蛋白表达的增强。结果还表明，该测试分子具有抑制 PTEN/PI3K/AKT 信号通路的潜力。
总之，本研究表明，橙皮素类黄酮有可能被开发为治疗胆囊腺癌的候选药物，但需要进行更多的毒理学和体内研究。
