Lupeol inhibits growth and migration in two human colorectal cancer cell lines by suppression of Wnt–β-catenin pathway

Abstract

BackgroundLupeol, a triterpene isolated from various herbal plants, possesses an anti-inflammatory function and has been proposed as a candidate for anticancer agents. The purpose of this research was to investigate the effect of lupeol on the viability, apoptosis, cell-cycle distribution, and migration of colorectal cancer cell lines and its molecular mechanism.MethodsLupeol was assessed for its anticancer effect using two human colorectal cancer cell lines: SW480 and HCT116. These cells were treated with lupeol, and their viability, apoptosis, migration, and cycle distribution were detected by CCK8, flow cytometry, and the transwell method. Quantitative PCR, Western blot, and immunofluorescence were applied to detect the expressions of CTNNB1, TCF4, cMYC, CCND1, CLDN1, and CCNA2.ResultsLupeol suppressed cell viability and migration and induced cellular apoptosis of both cell lines, with increased p53 and decreased Bcl2 protein levels (P<0.05). Cell cycles of both lupeol-treated cell lines were arrested in the S phase (P<0.05). Quantitative PCR and Western blot analyses showed significantly reduced expressions of CTNNB1, TCF4, and downstream genes of the Wnt–β-catenin pathway, including the cell-cycle-regulated genes of cMYC and CCND1 of both cell lines upon lupeol treatment (P<0.05). mRNA and protein levels of CLDN1 decreased in HCT116 cells, plus the expression of CCNA2 mRNA and protein decreased in SW480 cells (P<0.05). Immunofluorescence analysis confirmed decreased expression of Wnt–β-catenin signaling.ConclusionOur findings indicate that lupeol effectively inhibits proliferation and migration and induces apoptosis and cell-cycle arrest of two colorectal cell lines by inactivation of the Wnt–β-catenin signaling pathway and downregulation of cMYC, CCND1, CCNA2, and CLDN1, thereby making it a promising anticancer candidate.

Knockdown of KLK12 inhibits viability and induces�apoptosis in human colorectal cancer HT-29 cell line

“…MLK7-AS1 knockdown promoted CRC cell apoptosis in vitro (28). Lupeol-induced cellular apoptosis of both colorectal cancer cell lines, which increased p53 and decreased Bcl2 protein levels (29). In the present study, apoptosis levels and the expression levels of apoptosis-associated proteins were determined by flow cytometry and western blot analysis, respectively.…”

Determination of Lupeol, a Cytotoxic Compound Against SW620 Cells in the Extracts of Ha-Rak Recipe

“…13 According to the effective cytotoxic activities of lupeol against various cancer cells on multi-targeting pathways in previous reports, the compound has been suggested as a promising therapeutic and chemopreventive agent. [14][15][16][17] Thus, there is an evidence for supporting lupeol as a chemical marker agent in this continuing study. The chromatographic method using high-performance liquid chromatography (HPLC) with photodiode array detection (DAD) was considered to apply as an analytical method in our ongoing study, it was developed and validated for the analysis of lupeol in the extracts of 12 marketed Ha-Rak recipe.…”

Potential targeting of Hep3B liver cancer cells by lupeol isolated from Avicennia marina

“…[ 32 ] Lupeol also suppressed cell viability and migration and induced cellular apoptosis, as it also decreased protein expression of Wnt–β‐catenin pathway in colorectal cancer cells. [ 37 ] Therefore, lupeol can target multiple signaling pathways leading to induction of apoptosis and inhibition of growth. However, the anticancer activity of the compound, its efficacy, and mode of action on hepatocellular Hep3B have never been studied.…”