Kumar Prateek, Chauhan Anjali, Kumar Munendra, Kuanr Bijoy K, Kundu Aditi, Solanki Renu, Kapur Monisha Khanna
Microbial Technology Lab, Acharya Narendra Dev College, University of Delhi, Govindpuri, Kalkaji, New Delhi, 110 019 India.
School of Life Sciences, Jawaharlal Nehru University, New Delhi, 110067 India.
3 Biotech. 2021 Jun;11(6):254. doi: 10.1007/s13205-021-02812-w. Epub 2021 May 7.
During our previous investigation, bioactive compounds present in the extract of sp. strain 196 were characterized using LC-MS/MS and H NMR studies. These compounds were K-252-C aglycone indolocarbazole alkaloid, decoyinine, and cycloheximide; the study of these natural drugs against lung carcinoma is still limited. Focus of the current investigation was to study the anticancer effect of strain 196 extract on lung cancer cells (A549). During in vitro studies, anti-proliferative effect of extract was studied using MTT assay in A549 cells. Effect of extract on cell survival was further evaluated using colony assay. Cell death was qualitatively assessed using apoptosis assay. The aftereffect of extract treatment on metastatic potential of cancerous cells was studied using wound closure assay. Effect of extract on the morphology and cytoskeletal arrangement of A549 cells was studied using phalloidin staining. The extract demonstrated concentration and time-dependent cytotoxicity with IC value at 0.5 mg/ml (6 h) and 0.15 mg/ml (24 h). The proliferation and metastatic potential of cells, as characterized by MTT and migration assay, decreased over time in a concentration-dependent manner. Discrete changes in cellular morphology were noted as a result of the induced cytotoxicity. Apoptosis assay demonstrated 98.7% cell death at highest concentration of extract (1 mg/ml). During in silico studies, molecular docking revealed that strain 196 compounds are efficiently binding to mutant EGFR form (T790M/L858R) with release of binding energy (∆G) between - 5 and - 6.9 kcal/Mol. In conclusion, strain 196 extract could be a source of therapeutic drugs to treat lung carcinoma.
在我们之前的研究中，使用液相色谱-串联质谱法（LC-MS/MS）和核磁共振氢谱（¹H NMR）研究对sp.菌株196提取物中存在的生物活性化合物进行了表征。这些化合物是K-252-C苷元吲哚咔唑生物碱、脱氧精胍菌素和放线菌酮；这些天然药物对肺癌的研究仍然有限。当前研究的重点是研究菌株196提取物对肺癌细胞（A549）的抗癌作用。在体外研究中，使用MTT法研究了提取物在A549细胞中的抗增殖作用。使用集落形成试验进一步评估了提取物对细胞存活的影响。使用凋亡试验对细胞死亡进行了定性评估。使用伤口愈合试验研究了提取物处理对癌细胞转移潜能的后续影响。使用鬼笔环肽染色研究了提取物对A549细胞形态和细胞骨架排列的影响。提取物表现出浓度和时间依赖性细胞毒性，IC值在0.5 mg/ml（6小时）和0.15 mg/ml（24小时）。以MTT法和迁移试验表征的细胞增殖和转移潜能随时间呈浓度依赖性降低。由于诱导的细胞毒性，观察到细胞形态发生了明显变化。凋亡试验表明，在提取物最高浓度（1 mg/ml）下细胞死亡率为98.7%。在计算机模拟研究中，分子对接显示菌株196化合物与突变型表皮生长因子受体（EGFR）形式（T790M/L858R）有效结合，结合能释放（∆G）在-5至-6.9 kcal/mol之间。总之，菌株196提取物可能是治疗肺癌的治疗药物来源。
