Experimental study on inhibitory effects of diallyl sulfide on growth and invasion of human osteosarcoma MG-63 cells

Abstract

The inhibitory effects of diallyl sulfide (DAS) derived from allicin on in vitro and in vivo proliferation of human osteosarcoma MG-63 cells and the action mechanism, and the influence of DAS on invasive capability of MG-63 cells were investigated in order to search for the novel medicines for osteosarcoma. In the in vitro experiment, MG-63 cells were treated with different concentrations of DSA, and the morphological changes of MG-63 cells were observed under an inverted phase microscope. MTT method was used to assay the proliferation of MG-63 cells. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was used to detect the VEGF mRNA expression level in MG-63 cells. By using Transwell invasion assay, the influence of DAS on invasive ability of MG-63 cells was tested. In the in vivo experiment, the nude mice MG-63 cells tumor-bearing model was established, and different concentrations of DAS were injected beside the tumor. Twenty-one days after treatment, the mice were killed, the tumor size and tumor inhibition rate were calculated. The microvessel density (MVD) was determined by using immunohistochemistry. In the in vitro experiment, different concentrations of DAS could obviously inhibit proliferation of MG-63 cells in a time- and concentration-dependent manner. RT-PCR revealed that the expression levels of VEGF mRNA in DSA groups (different concentrations) were significant reduced as compared with those in control group (all P<0.05). Transwell invasion assay indicated that in 20 and 40 μg/mL DAS groups, the number of migratory cells was 91.4±8.3 and 81.8±7.4 respectively, which was significantly declined as compared with that in control group (150.4±14.7, both P<0.05). In the in vivo experiment, DAS could significantly suppress the growth of MG-63 tumor-bearing tissue. Immunohistochemistry demonstrated that different concentrations (20 and 40 μg/mL) of DAS could significantly decrease MVD of MG-63 tumor-bearing tissue (all P<0.05). It was suggested that DAS could inhibit the growth of MG-63 cells probably by suppressing the expression of VEGF mRNA.

Keywords

osteosarcoma / diallyl sulfide / growth inhibition / in vitro invasion

References

[1] ButtM.S., SultanM.T., ButtM.S., et al.. Garlic: nature’s protection against physiological threats. Crit Rev Food Sci Nutr, 2009, 49(6): 538-551

[2] MatsuuraN., MiyamaeY., YamaneK., et al.. Aged garlic extract inhibits angiogenesis and proliferation of colorectal carcinoma cells. J Nutr, 2006, 136(3): 842-846

[3] ParkS.Y., ChoS.J., KwonH.C., et al.. Caspase-independent cell death by allicin in human epithelial carcinoma cells: involvement of PKA. Cancer Lett, 2005, 224(1): 123-132

[4] LuH.F., SueC.C., YuC.S., ChenS.C., et al.. Diallyl disulfide (DADS) induced apoptosis undergo caspase-3 activity in human bladder cancer T24 cells. Food Chem Toxicol, 2004, 42(10): 1543-1552

[5] ChungJ.G., LuH.F., YehC.C., et al.. Inhibition of N-acetyltransferase activity and gene expression in human colon cancer cell lines by diallyl sulfide. Food Chem Toxicol, 2004, 42(2): 195-202

[6] HosonoT., FukaoT., OgiharaJ., et al.. Diallyl trisulfide suppresses the proliferation and induces apoptosis of human colon cancer cells through oxidative modification of beta-tubulin. J Biol Chem, 2005, 280(50): 41487-41493

[7] SakamotoK., LawsonL.D., MilnerJ.A.. Allyl sulfides from garlic suppress the in vitro proliferation of human A549 lung tumer cells. Nutr Cancer, 1997, 29(2): 152-156

[8] WuP.P., ChungH.W., LiuK.C., et al.. Diallyl sulfide induces cell cycle arrest and apoptosis in HeLa human cervical cancer cells through the p53-, caspase- and mitochondria-dependent pathways. Int J Oncol, 2011, 38(6): 1605-1613

[9] OommenS., AntoR.J., SrmivasG., et al.. Allicin (from garlic) induces caspase-mediated apoptosis in cancer cells. Eur J Pharmacol, 2004, 485(1–3): 97-103

[10] XiaoX.L., PengJ., SuQ., et al.. Diallyl trisulfide induces apoptosis of human gastric cancer cell line MGC803 through Caspase-3 pathway. Arch Pharm Res, 2006, 25(10): 1247-1251

[11] ArunkumarA., VijayababuM., GunadhariniN., et al.. Induction of apoptosis and histone hyperacetylation by diallyl disulfide in prostate cancer cell line PC-3. Cancer Lett, 2007, 251(1): 59-67

[12] DasA., BanikL., RayS.K.. Garlic compounds generate reactive oxygen species leading to activation of stress kinases and cysteine proteases for apoptosis in human glioblastoma T98G and U87MG cells. Cancer, 2007, 110(5): 1083-1095

[13] ZhangY.W., WenJ., XiaoJ.B., et al.. Induction of apoptosis and transient increase of phosphorylated MAPKs by diallyl disulfide treatment in human nasopharyngeal carcinoma CNE2 cells. Arch Pharm Res, 2006, 29(12): 1125-1131

[14] LachmannG., LorenzD., RadeckW., et al.. The pharmacokinetics of the S35 labeled labeled garlic constituents alliin, allicin and vinyldithiine. Arzneim-Forsch, 1994, 44(6): 734-743

[15] LedezmaE., Apitz-CastroR., CardierJ.. Apoptotic and anti-adhesion effect of ajoene, a garlic derived compound, on the murine melanoma B16F10 cells: possible role of caspase-3 and the alpha(4)beta(1) integrin. Cancer Lett, 2004, 206(1): 35-41

[16] BajpaiJ., SharmaM., SreenivasV., et al.. VEGF expression as a prognostic marker in osteosarcoma. Pediatr Blood Cancer, 2009, 53(6): 1035-1039

[17] YuanY.D., LiuQ.Y., YuanQ., et al.. Quantitative Study of Vascular Endothelial Growth Factor Expression in Malignant Bone Tumor. Chin J Orthop (Chinese), 1999, 19(7): 415-417

[18] ZengJ.X., HeX.S., MaY., et al.. Effect of allicin on the growth of hepatocellular carcinoma cells. Chin J Gen Surg (Chinese), 2009, 18(07): 691-694

[19] TakeiY., KadomatsuK., YuzawaY., et al.. A small interfering RNA targeting vascular endothelial growth factor as cancer therapeutics. Cancer Res, 2004, 64(10): 3365-3370

[20] FolkmanJ.. What is the evidence that tumors are angiogenesis dependent?. J Natl Cancer Inst, 1990, 82(1): 4-6