Anti-cancer activities of chalcone and flavanone derivatives in cancer cell line models

Abstract

In 2022, cervical cancer is the fourth most common cancer in female worldwide that causes human papillomaviruses (HPVs) infection. Nowadays, phytochemical compounds are used as anticancer agents and anticancer developments. In northern of Thailand, Syzygium nervosum A. Cunn. ex DC or Ma-kiang is a local plant which contains many biological activities such as anti-cancer, anti-oxidant and anti-viral activities. 2´,4´dihydroxy-6´-methoxy-3´,5´-dimethylchalcone (DMC) is a chalcone derivatives which is isolated from S. nervosum. In addition, the modification structure of parent phytochemical compounds could improve pharmacological activities such as anti-cancer activities by using chemical synthesis. The research aims to investigate the biological activities of DMC from S. nervosum A in human cervical cancer cell lines (C-33A, HeLa and SiHa). In addition, DMC and their derivatives (3a and 3b) were examined their biological activities for confirming the possible anti cervical cancer pathways by anti-migration assay, cell cycle assay, and mRNA expression analysis. As the results, DMC showed anti-proliferative activities in three human cervical cancer cell lines including C-33A, HeLa and SiHa, with IC50 values of 15.76 ± 1.49, 10.05 ± 0.22, and 18.31 ± 3.10 µM, respectively. Especially, DMC showed potential antiproliferative cervical cancer activities in HeLa cells. DMC-induced apoptosis cell death in HeLa cells was then investigated by DNA damage, cell cycle arrest, and apoptosis assays. DMC treatment increased DNA damage in HeLa cells, observed by fluorescence inverted microscopy and a comet assay. The cell cycle assays increased G0/G1 phases after treatment with DMC indicated G0/G1 cell cycle arrest. Afterward, DMC treatment-induced apoptosis cell death in HeLa cells was approximately three- to four-fold higher compared with the untreated group. These findings showed that DMC a phytochemical compound, is a potential anti-cervical cancer agent and anti-cancer development. Subsequently, DMC and their derivatives (3a and 3b) showed potential antiproliferative with IC50 values of 7.56 ± 0.27 and 8.24 ± 0.14 µM, respectively in SiHa cells, which were approximately two-fold greater than DMC. Next, compound 3a and 3b slightly inhibited SiHa cell migration in the wound healing assay. The cell cycle analysis of SiHa cells after treatment with compound 3a and 3b increased G0/G1 phases that indicated inhibition of cell cycle. Moreover, compound 3a showed potential anti-cancer activities with upregulating TP53 and CDKN1A which resulted in upregulation of BAX and downregulation of CDK2 and BCL2, leading to cell cycle arrest and apoptosis. The BAX/BCL2 expression ratio was increased after treatment with compound 3a in SiHa cells, leading to the intrinsic apoptotic pathway. In silico molecular dynamics simulation investigated on how these DMC derivatives interact with the HPV16 E6 protein, a viral oncoprotein associated with cervical cancer. Therefore, the findings suggest that DMC and their derivative (3a) an potential candidate for anti-cervical cancer drugs. The information will be beneficial to a better understanding in the anti-cancer pathway and anti-cancer drug development.