Selective AhR receptor modulator 3,3′-Diindolylmethane (DIM) is a class of relatively non-toxic indole derivatives. DIM is an acid-catalyed consendation product of indole-3-carbinol, a consititudent of cruciferous vegetables, and is formed in the stomach [12]. DIM is an anti-cancer agent, it suppresses cancer cell proliferation in mammary [13], colon [14] and pancreatic [15] cancers. There had been little reports about the effects of DIM on gastric cancer cells growth, the present study was designed to observe

the effects of DIM on gastric cancer cells growth and explore the possible mechanisms. Methods Cell line Human gastric cancer cell line SGC7901 was obtained from the buy AZD1208 Cancer Institute of Chinese Academy Ipatasertib purchase of Medical Science. SGC7901 Cells were maintained in RPMI-1640 medium (GIBCO, Carlsbad, Calif, USA) supplemented with 10% fetal bovine serum (Hyclone, USA), 1 × 105 U/L of penicillin, and 0.1 g/L of gentamycin. The cellular environment was maintained at 50 mL/L CO2 and 37°C. Treatment of cells DIM was purchased from Enzo Life Science company (Bulter Pike plymouth meeting, PA, USA), resveratrol and dimethyl sulfoxide (DMSO) were purchased from Sigma Chemical Company (Bellefonte, PA, USA). DIM and resveratrol were dissolved in DMSO. After incubating for 24 h, one group of cells was treated with DIM at different

concentrations (0, 10, 20, 30, 40, 50 μmol/L) for 24 hours. A second group was treated with DIM (30 μmol/L) plus resveratrol (0, 1, 5, 10, 20 μmol/L) for

6 h. Another group was treated with DIM (30 μmol/L) for different time intervals (0, 1, 6, 24, 48, 72 h), respectively. Control cells received 1 mL/L DMSO only. Reverse transcription–polymerase chain reaction (RT-PCR) After harvesting the cell, total RNA was extracted using the Qiagen RNeasy Mini Kit (Qiagen, Germany) according to the manufacturer’s instructions. cDNA was synthesized with 1 μg total RNA using reverse transcriptase, Phosphoglycerate kinase ReverTraAceTM (Toyobo Co., Osaka, Japan) under the following conditions: 30°C for 10 min, 42°C for 20 min, 99°C for 5 min, and 4°C for 5 min. Polymerase chain reaction (PCR) was performed using 2 μl of complementary DNA and 0.6 U Ex Taq DNA polymerase (Takara, Dalian, China ) in 20 μl reaction system and for 30 cycle with 94°C denaturation for 30 s, 55°C annealing for 30 s and 72°C elongation for 45 s. The primer sequences were as follows: reverse transcription–polymerase chain reaction (RT–PCR): AhR, 5’- ACT CCA CTT CAG CCA CCA TC -3’ (forward) and 5’- ATG GGA CTC GGC ACA ATA AA -3’ (reverse), the proposed size of PCR product was 204 bp. CYP1A1, 5’- CCA TGT CGG CCA CGG AGT T -3’(forward) and 5’- ACA GTG CCA GGT GCG GGT T -3’ (reverse), the proposedsize of PCR product was 174 bp.