Allyl Isothiocyanate Inhibits the Proliferation of Renal Carcinoma Cell Line GRC-1 by Inducing an Imbalance Between Bcl2 and Bax

Abstract

BackgroundBecause of the insensitivity of renal cell carcinoma (RCC) to both chemotherapy and radiotherapy, surgery remains the primary approach for anticancer treatment. However, patients who do not receive timely diagnoses may not be suitable for surgery, especially in the late phase of tumor development. Thus, the discovery of novel effective treatment is of great importance. Allyl isothiocyanate (AITC) can inhibit the proliferation and induce apoptosis in many cancer cells. In this paper, we report on an in vitro study to determine the effect of AITC on proliferation and apoptosis of RCC line GRC-1.Material/MethodsCCK8 assay was used to detect cell proliferation under gradient concentrations of AITC. Flow cytometry was employed to evaluate cell apoptosis. Real-time fluorescent polymerase chain reaction quantified mRNA levels of Bax and Bcl-2 genes. Western blotting was further employed for protein expression assay.ResultsAITC inhibited GRC-1 cell proliferation and induced cell apoptosis in a dose-dependent manner; it also elevated Bax while suppressing Bcl-2 gene expression at both mRNA and protein levels. In general, increasing concentration of AITC decreased Bcl-2/Bax ratio.ConclusionsThe inhibitory effect of AITC on GRC-1 cells is exerted via cell apoptosis, in which the imbalance of Bcl-2/Bax plays a significant role.

Expression and epigenetic regulatory mechanism of BNIP3 in clear cell renal cell carcinoma

“…DNA methylation is a proven mechanism of BNIP3 downregulation in tumors. In a previous study, Murai and coworkers demonstrated that BNIP3 was methylated in 65.6% of colorectal cancer tissues, although it was not methylated in adjacent normal tissue samples ( 15 , 40 ). Similarly, Cleven et al ( 41 ) also demonstrated the occurrence of BNIP3 methylation in 52.8% of colorectal cancer cells, and that treatment with 5-aza-C restored the expression of BNIP3 and led to increased apoptosis and autophagy, with enhanced sensitivity to chemotherapy.…”

“…Following cell dissociation and propagation, the 786-O, A498, ACHN, and GRC-1-1 cell lines were cultured (37°C) and grown in Roswell Park Memorial Institute (RPMI) medium using 1640 complete medium (Gibco ® ; Thermo Fisher Scientific, Inc., Waltham, MA, USA). The GRC-1 RCC line was established at the Institute of Urology, Peking University (Beijing, China), was first reported by Ding et al ( 14 ), and has been subsequently used in numerous studies ( 15 , 16 ). PC3 and Du145 cells were cultured (37°C) in Dulbecco’s modified Eagle’s medium (DMEM) complete medium (Gibco ® ; Thermo Fisher Scientific, Inc.), whereas HK-2 cells were cultured (37°C) in F-12 Complete™ medium (Gibco ® ; Thermo Fisher Scientific, Inc.) in microcentrifuge tubes (Eppendorf, Stevenage, UK) in a humidified incubator in an atmosphere of 5% CO 2 and 95% air.…”

Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells

“…Moreover, blocking Ca 2+ -based ER stress signals and ROS formation significantly attenuated AITC-induced HT-29 cell apoptosis. The effective cytotoxic actions of AITC on multiple human cancer cell lines are well documented (12,(16)(17)(18)(19)(20)(21); however, the detailed mechanisms involved in the cytotoxic effects of AITC in HT-29 cells remain elusive. Our results indicated that AITC inhibited HT-29 cell growth and proliferation.…”

Overexpression of ATP citrate lyase in renal cell carcinoma tissues and its effect on the human renal carcinoma cells in�vitro

“…Human RCC, 786-0 and GRC-1 ( 7 ), and normal human kidney cells, HK-2 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). All of the cells were maintained in DMEM (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) and incubated at 37°C in humidified air with 5% CO 2 .…”