Sibuh Belay Zeleke, Khanna Sonia, Taneja Pankaj, Sarkar Paratpar, Taneja Neetu Kumra
Department of Biotechnology, School of Engineering and Technology, Sharda University, Greater Noida, U. P., India.
Department of Chemistry and Biochemistry, School of Basic Sciences and Research, Sharda University, Greater Noida, U. P., India.
Life Sci. 2021 May 15;273:119305. doi: 10.1016/j.lfs.2021.119305. Epub 2021 Mar 3.
The aim of this study was to synthesize and evaluate anticancer activity of 2-hydroxy benzaldehyde and 4-hydroxy benzaldehyde thiosemicarbazone (2-HBTSc and 4-HBTSc) against MCF-7 breast cancer cell line.
The ligands were prepared and characterized by UV vis, IR and NMR. MTT assay was used to assess viability of cells. RNA isolation, extraction and cDNA synthesis were done. Then all groups were subjected to RT-qPCR using Gene expression specific primers. Also, western blot protein expression and molecular docking were done. Two-way ANOVA with Tukey post-hoc test was employed to test the significance using GraphPad Prism.
The IC values were 3.36μg/ml and 3.60μg/ml for 2-HBTSc and 4-HBTSc treated MCF-7 tumor cells respectively. Tumor cell growth inhibition ranged from 38 to 49.27% in 4-HBTSc treated cells, and 19 to 25% in 2-HBTSc treated cells with increase in doses 5 μg/ml to 20 μg/ml. The protein and gene expression result showed a significant upregulation in tumor suppressor and apoptosis inducing genes while, oncogene activity was significantly downregulated. Specifically, BRCA2 and pRB gene showed the highest expression in 4-HBTSc and 2-HBTSc treated cells respectively. Conversely, RAS oncogene was downregulated significantly. Docking result showed that both 2-HBTSc and 4-HBTSc have the potential to inhibit Estrogen Receptor Alpha Ligand Binding Domain, Human 17-Beta-hydroxysteroid dehydrogenase type 1 mutant protein and Human Topoisomerase II alpha that are expressed more during Breast Cancer.
The findings of this study imply that the test compound has potential for further study.
本研究旨在合成并评估2-羟基苯甲醛和4-羟基苯甲醛缩氨基硫脲（2-HBTSc和4-HBTSc）对MCF-7乳腺癌细胞系的抗癌活性。
通过紫外可见光谱、红外光谱和核磁共振对配体进行制备与表征。采用MTT法评估细胞活力。进行RNA分离、提取和cDNA合成。然后使用基因表达特异性引物对所有组进行RT-qPCR。同时进行蛋白质印迹蛋白表达和分子对接。使用GraphPad Prism软件采用双向方差分析及Tukey事后检验来检验显著性。
2-HBTSc和4-HBTSc处理MCF-7肿瘤细胞的IC值分别为3.36μg/ml和3.60μg/ml。随着剂量从5μg/ml增加到20μg/ml，4-HBTSc处理组肿瘤细胞生长抑制率在38%至49.27%之间，2-HBTSc处理组在19%至25%之间。蛋白质和基因表达结果显示，肿瘤抑制和凋亡诱导基因显著上调，而癌基因活性显著下调。具体而言，BRCA2和pRB基因分别在4-HBTSc和2-HBTSc处理的细胞中表达最高。相反，RAS癌基因显著下调。对接结果表明，2-HBTSc和4-HBTSc均有可能抑制雌激素受体α配体结合域、人17-β-羟基类固醇脱氢酶1型突变蛋白和人拓扑异构酶IIα，这些蛋白在乳腺癌期间表达更多。
本研究结果表明受试化合物具有进一步研究潜力。
