评价提取物对人肝癌 HepG2 细胞和正常 HEK293T 细胞系的细胞毒性。

Evaluation of Cytotoxic Potential of Extracts on Human Hepatocarcinoma HepG2 and Normal HEK293T Cell Lines.

机构信息

Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore 54000, Pakistan.

Institute of Biochemistry and Biotechnology, University of Veterinary and Animal Sciences, Lahore 54000, Pakistan.

出版信息

Biomed Res Int. 2022 Sep 22;2022:1279961. doi: 10.1155/2022/1279961. eCollection 2022.

DOI:10.1155/2022/1279961

PMID:36193312

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9526597/

Abstract

Data regarding the therapeutic potential of () are insufficient. It becomes more important to explore plants as an alternative or palliative therapeutics in deadly diseases around the globe. The current study was planned to explore for its anticancer activity of ethanolic and hexane extracts of leaves against hepatic carcinoma (HepG2) and human embryonic kidney (HEK293T) cell lines. HepG2 and HEK293T cells were treated with 10, 50, 100, 200, and 400 g/mL of ethanolic and hexane extracts of and were incubated for 72 h. Antiproliferative activity was measured by 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assay, and percentage viability were calculated through crystal violet staining and cellular morphology by Floid Cell Imaging Station. The study showed ethanolic extract exhibiting a significantly higher antiproliferative effect on HepG2 (IC50 = 31g/mL) in a concentration-dependent manner, while HEK293T (IC50 = 241g/mL) cells showed no toxicity. Hexane extract exhibited lower cytotoxicity (IC50 = 150g/mL) on HepG2 cells with no effect on HEK293T (IC50 = 550g/mL). On the other hand, the percentage viability of HepG2 cells was recorded as 78%, 67%, 50%, 37%, and 28% by ethanolic extracts, and 88%, 80%, 69%, 59%, and 50% by hexane extracts at tested concentrations of both extracts. Toxicity assay showed significantly safer ranges of percentage viabilities in normal cells (HEK293T), i.e., 95%, 90%, 88%, 76%, and 61% with ethanolic extract and 97%, 95%, 88%, 75%, and 62% with hexane extract. The assay validity revealed 100% viability in the control negative (dimethyl sulfoxide treated) and less than 45% in the control positive (cisplatin) on both HepG2 and HEK293T cells. Morphological studies showed alterations in HepG2 cells upon exposure to >50 g/mL of ethanolic extracts and ≥400 g/mL of hexane extracts. HEK293T on the other hand did not change its morphology against any of the extracts compared to the aggressive changes on the HepG2 cell line by both extracts and positive control (cisplatin). In conclusion, extracts of are proved to have significant potential for cytotoxicity-induced apoptosis in human cancer HepG2 cells and are less toxic to normal HEK293T cells. Hence extracts are proposed to be used against hepatocellular carcinoma (HCC) after further validations.

摘要

关于 () 的治疗潜力的数据不足。在全球范围内，探索植物作为治疗致命疾病的替代或姑息疗法变得更加重要。本研究旨在探索 () 的叶乙醇和己烷提取物对肝癌 (HepG2) 和人胚肾 (HEK293T) 细胞系的抗癌活性。用 10、50、100、200 和 400μg/mL 的乙醇和己烷提取物处理 HepG2 和 HEK293T 细胞，并孵育 72h。通过 3-(4,5-二甲基噻唑-2-基)-2,5-联苯四唑溴盐 (MTT) 测定法测量增殖活性，并通过结晶紫染色和 Floid Cell Imaging Station 中的细胞形态计算细胞活力百分比。研究表明，乙醇提取物对 HepG2 表现出显著更高的浓度依赖性增殖抑制作用（IC50 = 31μg/mL），而 HEK293T（IC50 = 241μg/mL）细胞无毒性。己烷提取物对 HepG2 细胞的细胞毒性较低（IC50 = 150μg/mL），对 HEK293T（IC50 = 550μg/mL）无影响。另一方面，乙醇提取物处理的 HepG2 细胞的存活率百分比分别为 78%、67%、50%、37%和 28%，而己烷提取物的存活率百分比分别为 88%、80%、69%、59%和 50%在测试浓度下的两种提取物。毒性测定显示，正常细胞（HEK293T）的存活率百分比具有明显更安全的范围，即乙醇提取物为 95%、90%、88%、76%和 61%，己烷提取物为 97%、95%、88%、75%和 62%。该测定的有效性显示，在 HepG2 和 HEK293T 细胞上，阴性对照（用二甲基亚砜处理）的活力为 100%，阳性对照（顺铂）的活力小于 45%。形态学研究表明，暴露于 >50μg/mL 的乙醇提取物和 ≥400μg/mL 的己烷提取物后，HepG2 细胞的形态发生改变。另一方面，与两种提取物和阳性对照（顺铂）对 HepG2 细胞系的剧烈变化相比，HEK293T 细胞的形态没有变化。总之，提取物被证明在人类癌症 HepG2 细胞中具有显著的细胞毒性诱导细胞凋亡的潜力，对正常 HEK293T 细胞的毒性较小。因此，建议在进一步验证后将提取物用于治疗肝细胞癌 (HCC)。