背景:印度东北部有巨大的治疗性植物区系,Otteliaalismoides是一种水生植物,长期以来一直在传统医学中用于治疗癌症等疾病,结核病,糖尿病,Febrifuge,痔疮,和rubefacient。在肺癌和皮肤癌细胞中具有高的增殖和转移率,包括耐药性和非特异性靶标活性,对他们的治疗策略产生了重要的挑战。因此,寻找新的治疗靶点来治疗肺癌和皮肤癌的进展对提高患者的生存率至关重要。
目的:本研究的目的是评估O.alismoides的丙酮提取物的凋亡潜能(L.)Pers。(OA-AC)并确定导致这种效应的化合物,已经进行了提取物的HRLC-MS-QTOF分析以及鉴定的化合物的计算机内分子对接分析。
方法:A549和A431细胞在暴露24h和48h时用O.alismoides的丙酮提取物(OA-AC)处理,评估细胞周期阶段分布,并通过OA-EtBr染色和线粒体外膜电位测定评估凋亡诱导活性。进行Western印迹以评估凋亡蛋白表达。最后,分析OA-AC的HR-LCMS以鉴定负责提取物凋亡活性的化合物。
结果:在暴露于10μg/mL和25μg/mL的OA-AC24小时时,A549和A431细胞中的细胞周期阶段分布分析显示在细胞周期的G2/M期有效的阻滞或阻断,细胞周期蛋白B和p-Cdc2的表达降低。在暴露48小时时,在这些癌细胞中观察到凋亡,Bax的表达升高,p21和裂解的半胱天冬酶3并降低Bcl2的表达。
结论:这些癌细胞的AO-EtBr染色显示,OA-AC诱导的死亡本质上是凋亡性的,线粒体膜由于线粒体膜的丢失或损伤而去极化。OA-AC的HRLC-MS-QTOF分析描绘了14种主要的可分离化合物,分子对接分析显示了4种化合物,其可能作为细胞周期蛋白B的抑制剂用于G2/M期停滞,导致细胞中的凋亡诱导。
BACKGROUND: The North-eastern parts of India have immense therapeutic floras, Ottelia alismoides is an aquatic plant that has been in use for a long time in traditional medicine for treating diseases like cancer, tuberculosis, diabetes, febrifuge, hemorrhoids, and rubefacient. In lung and skin carcinoma cells with a high rate of proliferation and metastasis including drug resistance and non-specific target activity, generates important challenges towards their treatment strategy. Thus, finding novel therapeutic targets to treat lung and skin cancer progression is essential to enhance the patients\' survival with treatment.
OBJECTIVE: The purpose of this study was to evaluate the apoptotic potential of acetone extract of O. alismoides (L.) Pers. (OA-AC) and to identify the compounds responsible for this effect, HRLC-MS-QTOF analysis of the extract has been undertaken along with in-silico molecular docking analysis of the identified compounds.
METHODS: A549 and A431 cells were treated with acetone extract of O. alismoides (OA-AC) at 24 h and 48 h exposure and cell cycle phase distribution was evaluated and also apoptosis induction activity was evaluated by OA-EtBr staining and Mitochondrial outer membrane potential assay. Western blotting was performed for the evaluation of apoptotic protein expression. At last, the HR-LCMS of OA-AC was analyzed to identify the compounds responsible for the apoptotic activity of the extract.
RESULTS: The cell cycle phase distribution analysis in A549 and A431 cells at 24hrs exposure with 10 μg/mL and 25 μg/mL of OA-AC showed a potent arrest or blockage at the G2/M phase of the cell cycle with reduced expression of cyclin B and p-Cdc2. At 48 h exposure, apoptosis was observed in these cancer cells with elevated expression of Bax, p21 and cleaved caspase 3 and reduced expression of the Bcl2.
CONCLUSIONS: AO-EtBr staining of these cancer cells reveals that the death induced by OA-AC was apoptotic in nature with depolarization of mitochondrial membrane due to loss or damage of the mitochondrial membrane. The HRLC-MS-QTOF analysis of OA-AC depicted 14 major isolable compounds and molecular docking analysis displayed 4 compounds that might act as an inhibitor of cyclin B for G2/M phase arrest that leads to apoptotic induction in the cells.