Abstract
Achyranthes aspera (AA) has been used traditionally for the cure of various disorders. However, the action of root extracts of AA as anticancer agent and its cellular mechanism remain unclear. The aim was to screen the antitumor effect of ethanolic (EAA) and aqueous (AAA) root extracts on the growth of colon cancer COLO-205 cells by testing their cytotoxicity, followed by their effect on clonogenicity, migration, and induction of apoptosis. Mechanisms leading to apoptosis and cell cycle arrest were also investigated by expression studies of caspase-9, caspase-3, Bax, Bcl-2, p16, p21, and p27 genes, followed by flow cytometric analysis for cell cycle distribution. Cytotoxicity screening of AA extracts indicated greater cytotoxic activity of AAA extract against COLO-205 cells. A series of events marked by apoptosis revealed loss of cell viability, chromatin condensation, and DNA fragmentation in AAA treated cells to a greater extent. The mRNA expression levels of caspase-9, caspase-3, Bax, p16, p21, and p27 were markedly increased in the AAA treated cells, along with decreased Bcl-2 expression. The cell cycle arrest at S phase was detected by flow cytometric analysis after treatment with AAA. Overall the study signifies the aqueous extracts as a promising therapeutic candidate against cancer.
Highlights
Despite significant advances toward targeted therapy and screening techniques, colon cancer continues to be a chronic disease worldwide, being the third leading cause of death in men and the second in women globally
The results showed that Extract of Achyranthes aspera Roots (EAA) contained greater amounts of phenolic compounds with a TPC value of 419.2 ± 1.3 μg gallic acid equivalents (GAE)/gram of dry extract as compared to Achyranthes aspera Roots (AAA) having 273.8 ± 2.5 μg GAE/gram of dry extract of total phenolic content
The results showed EAA to exhibit greater antioxidant activity with a FRAP value of 195 ± 1.2 μM Fe (II)/gram of dry extract and IC50 value of DPPH radical of 288.3 ± 1.4 μg/mL of dry extract as compared to AAA with FRAP value of 121±1.5 and IC50 value of DPPH radical 372.6 ± 1.7
Summary
Despite significant advances toward targeted therapy and screening techniques, colon cancer continues to be a chronic disease worldwide, being the third leading cause of death in men and the second in women globally. According to the Globocan 2012 Cancer Fact Sheet, about 1.36 million new cases of colon cancer were clinically diagnosed, with number of deaths being 0.69 million [1]. In the development of cancer, evasion of apoptosis is one of the major factors resulting in overpopulation of cancer cells. Apoptosis is an active form of cell death guided by a set of prosurvival and antisurvival genes [2]. There is a strong corelation between loss of apoptotic control and cancer initiation and progression, as tumor cells lose their ability to activate the death signalling pathway [3]. Other than apoptosis, deregulated cell-cycle control is a key feature of cancer progression. The cell cycle begins or stops only in response to proliferationenhancing or retarding signals, respectively, which is not seen in cancer cells. As a result of this, their proliferation remains unchecked [4]
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