Sengupta Pracheta, Raman Sukanya, Chowdhury Rajdeep, Lohitesh K, Saini Heena, Mukherjee Sudeshna, Paul Atish
Department of Pharmacy, Birla Institute of Technology and Science, Pilani, India.
Department of Biological Sciences, Birla Institute of Technology and Science, Pilani, India.
Front Oncol. 2017 Dec 11;7:296. doi: 10.3389/fonc.2017.00296. eCollection 2017.
Cancer is a multifactorial disease and hence can be effectively overcome by a multi-constituently therapeutic strategy. Medicinal plant extracts represent a perfect example of such stratagem. However, minimal studies have been done till date that portray the effect of extraction techniques on the phyto-constituent profile of plant extracts and its impact on anticancer activity. In the present study, we have evaluated the anticancer potential of methanolic extracts of root and seeds prepared by various extraction techniques in human osteosarcoma (HOS) cells. Soxhlation extract of (BAM-SX) and sonication extract of (AIM-SO) were most effective in inducing apoptosis in parental drug sensitive, as well as resistant cell type developed by repeated drug exposure. Generation of reactive oxygen species and cell cycle arrest preceded caspase-mediated apoptosis in HOS cells. Interestingly, inhibition of autophagy enhanced cell death suggesting the cytoprotective role of autophagy. Combination studies of different methanolic extracts of BAM and AIM were performed, among which, the combination of BAM-SO and AIM-SO (BAAISO) was found to show synergism (IC 10.27 µg/ml) followed by combination of BAM-MC and AIM-MC (BAAIMC) with respect to other combinations in the ratio of 1:1. BAAISO also showed synergism when it was added to cisplatin-resistant HOS cells (HCR). Chromatographic profiling of BAM-SX and AIM-SO by high performance thin layer chromatography resulted in identification of berberine (R 0.55), palmitine (R 0.50) in BAM-SX and azadirachtin A (R 0.36), azadirachtin B (R 0.56), nimbin (R 0.80), and nimbolide (R 0.43) in AIM-SO. The cytotoxic sensitivity obtained can be attributed to the above compounds. Our results highlight the importance of extraction technique and subsequent mechanism of action of multi-constituential and against both sensitive and drug refractory HOS cells.
癌症是一种多因素疾病，因此可以通过多成分治疗策略有效攻克。药用植物提取物就是这种策略的一个完美范例。然而，迄今为止，关于提取技术对植物提取物中植物成分谱的影响及其对抗癌活性的影响的研究极少。在本研究中，我们评估了通过各种提取技术制备的根和种子的甲醇提取物对人骨肉瘤（HOS）细胞的抗癌潜力。（BAM-SX）的索氏提取提取物和（AIM-SO）的超声提取提取物在诱导亲代药物敏感细胞以及通过反复药物暴露产生的耐药细胞类型凋亡方面最为有效。活性氧的产生和细胞周期阻滞先于HOS细胞中半胱天冬酶介导的凋亡。有趣的是，自噬的抑制增强了细胞死亡，表明自噬具有细胞保护作用。对BAM和AIM的不同甲醇提取物进行了联合研究，其中，发现BAM-SO和AIM-SO的组合（BAAISO）显示出协同作用（IC 10.27μg/ml），其次是BAM-MC和AIM-MC的组合（BAAIMC），相对于其他比例为1:1的组合。当将BAAISO添加到顺铂耐药的HOS细胞（HCR）中时，也显示出协同作用。通过高效薄层色谱对BAM-SX和AIM-SO进行色谱分析，结果在BAM-SX中鉴定出小檗碱（R 0.55）、棕榈碱（R 0.50），在AIM-SO中鉴定出印楝素A（R 0.36）、印楝素B（R 0.56）、尼姆宾（R 0.80）和尼姆比德（R 0.43）。所获得的细胞毒性敏感性可归因于上述化合物。我们的结果突出了提取技术以及多成分提取物对敏感和耐药HOS细胞的后续作用机制的重要性。
