Gunathilaka Thilina Lakmini, Dilrangi Kulathungage Hiranthi, Ranasinghe Pathmasiri, Samarakoon Kalpa W, Peiris L Dinithi C
Department of Zoology, Faculty of Applied Sciences, University of Sri Jayewardenepura, Nugegoda 10250, Sri Lanka.
Industrial Technology Institute, Halbarawa Gardens, Malabe 10115, Sri Lanka.
Pharmaceuticals (Basel). 2021 Nov 12;14(11):1154. doi: 10.3390/ph14111154.
The current study determined the cytotoxic and apoptotic potential of the polyphenol-rich methanol extract of () and its fractions against human breast adenocarcinoma (MCF-7) and rhabdomyosarcoma (RMS) cells. MTT and neutral red assays were used to determine cytotoxicity. The clonogenic assay evaluated the antineoplastic activity, while the apoptotic activity was determined by cellular morphological changes, caspase 3/7 activity, and DNA fragmentation. Morphological alterations in apoptosis were observed by an inverted phase-contrast microscope and Hoechst 33342 staining methods. The total phenolic, flavonoids, alkaloids, and antioxidant activity in the hexane and chloroform fractions were determined, based on their cytotoxic activity. The hexane fraction of effectively reduced the cell growth that is concentration-dependent in human RMS and MCF-7 cell lines. It also exhibited low cytotoxicity on Vero cells. The characteristic cellular and nuclear apoptotic morphological features were observed. A noticeable caspase 3/7 activation and the fragmented DNA were detected only in the hexane fraction treated RMS cells, whereas MCF-7 cells showed low caspase 3/7 activation due to a lack of caspase 3 and no evidence of having a typical ladder pattern of apoptosis. Further analysis revealed that the hexane fraction-treated RMS cells upregulated the gene twofold (2.72) compared to the (0.77) gene, whereas in the MCF-7 cells, a 2.21-fold upregulation of was observed compared to the (0.64) gene. The hexane fraction exhibited moderate total phenolics, flavonoids, alkaloids content, and antioxidant activity. According to the different antioxidant mechanisms, hexane and chloroform fractions showed the highest antioxidant activities by FRAP and ORAC assays, respectively. GC-MS analysis of hexane fraction revealed the presence of methyl tetradecanoate (38.314%) as the most abundant compound. The study's findings highlighted that the non-polar compounds present in the hexane fraction of suppressed cell proliferation and induced apoptosis-mediated cell death in RMS and MCF-7 cells, mainly via the activation of the gene. Hence, the isolation of compounds is warranted. However, more studies are required to understand the mechanistic insights of these observations.
本研究确定了[植物名称]富含多酚的甲醇提取物及其馏分对人乳腺腺癌（MCF - 7）和横纹肌肉瘤（RMS）细胞的细胞毒性和凋亡潜力。采用MTT法和中性红法测定细胞毒性。克隆形成试验评估抗肿瘤活性，而凋亡活性则通过细胞形态变化、半胱天冬酶3/7活性和DNA片段化来确定。通过倒置相差显微镜和Hoechst 33342染色法观察凋亡过程中的形态学改变。根据己烷和氯仿馏分的细胞毒性活性，测定了其中的总酚、黄酮、生物碱和抗氧化活性。[植物名称]的己烷馏分有效地抑制了人RMS和MCF - 7细胞系中浓度依赖性的细胞生长。它对Vero细胞也表现出低细胞毒性。观察到了典型的细胞和细胞核凋亡形态特征。仅在经己烷馏分处理的RMS细胞中检测到明显的半胱天冬酶3/7激活和DNA片段化，而MCF - 7细胞由于缺乏半胱天冬酶3，半胱天冬酶3/7激活程度较低，且没有典型的凋亡梯状模式的证据。进一步分析表明，与[对照基因]（0.77）相比，经己烷馏分处理的RMS细胞中[目标基因]上调了两倍（2.72），而在MCF - 7细胞中，与[对照基因]（0.64）相比，[目标基因]上调了2.21倍。己烷馏分表现出中等的总酚、黄酮、生物碱含量和抗氧化活性。根据不同的抗氧化机制，己烷和氯仿馏分分别通过FRAP和ORAC试验表现出最高的抗氧化活性。己烷馏分的GC - MS分析表明，十四烷酸甲酯（38.314%）是最主要的化合物。该研究结果突出表明，[植物名称]己烷馏分中的非极性化合物主要通过激活[目标基因]抑制RMS和MCF - 7细胞的增殖并诱导凋亡介导的细胞死亡。因此，有必要对化合物进行分离。然而，需要更多的研究来了解这些观察结果的机制。
