Ganoderma lucidum inhibits proliferation of human ovarian cancer cells by suppressing VEGF expression and up-regulating the expression of connexin 43

BMC Complementary and Alternative Medicine

Ning Wang¹  Xiaofei Sun¹  Jingjing Liu¹  Shuyan Dai¹ 

[1] Department of Obstetrics and Gynecology, The Affiliated Shengjing Hospital, China Medical University, 36 Sanhao Street, Heping district, Shenyang 110004, People’s Republic of China

关键词: Connexin 43 (Cx43);    Vascular endothelial growth factor (VEGF);    Immunohistochemistry;    Real-time PCR;    Ovarian cancer;    Ganoderma lucidum;   

Others  :  1085606
DOI  :  10.1186/1472-6882-14-434

 received in 2013-10-01, accepted in 2014-10-27,  发布年份 2014

【 摘 要 】

Background

Ganoderma lucidum (G. lucidum, Reishimax) is an herbal mushroom known to have inhibitory effect on tumor cell growth. However, the molecular mechanisms responsible for its anti-proliferative effects on the ovarian cancer have not been fully elucidated.

Methods

Human ovarian cancer cells HO 8910 (HOCC) and human primary ovarian cells (HPOC) were treated with G. lucidum. Effects of G. lucidum treatment on cell proliferation were studied by MTT assay. The expression of vascular endothelial growth factor (VEGF) and connexin 43 (Cx43) were measured by immunohistochemistry and real time polymerase chain reaction. To study the molecular mechanism of CX43 mediated anti-tumor activity, small interference RNA (siRNA) was used to knockdown Cx43 expression in HOCC.

Results

G. lucidum treatment resulted in reduced proliferation of HOCC. Inhibition of proliferation was accompanied by a decrease in VEGF expression and increase in Cx43 expression in the cancer cells. The extent of immune-reactivity of Cx43 or VEGF in cancer cells were correlated with the concentrations of G. lucidum used for treatment. Furthermore, knockdown of Cx43 expression in HOCC abrogated the effect of G. lucidum on cell proliferation without alteration of G. lucidum-induced attenuation of VEGF expression.

Conclusions

G. lucidum inhibits ovarian cancer by down-regulating the expression of VEGF and up-regulating the downstream Cx43 expression. G. lucidum may be a promising therapeutic agent for the treatment of ovarian cancer.

【 授权许可】

   
2014 Dai et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin 2011, 61(2):69-90.
• [2]Wu GS, Guo JJ, Bao JL, Li XW, Chen XP, Lu JJ, Wang YT: Anti-cancer properties of triterpenoids isolated from Ganoderma lucidum - a review. Expert Opin Investig Drugs 2013, 22(8):981-992.
• [3]Hsieh T-C, Wu JM: Suppression of proliferation and oxidative stress 1 by extracts of Ganoderma lucidum in the ovarian cancer cell line OVCAR-3. Int J Mol Med 2011, 28(6):1065-1069.
• [4]Zhao S, Ye G, Fu G, Cheng J-X, Yang BB, Peng C: Ganoderma lucidum exerts anti-tumor effects on ovarian cancer cells and enhances their sensitivity to cisplatin. Int J Oncol 2011, 38(5):1319-1327.
• [5]Santin A, Hermonat P, Ravaggi A, Cannon M, Pecorelli S, Parham G: Secretion of vascular endothelial growth factor in ovarian cancer. Eur J Gynaecol Oncol 1999, 20(3):177-181.
• [6]Mesiano S, Ferrara N, Jaffe RB: Role of vascular endothelial growth factor in ovarian cancer: inhibition of ascites formation by immunoneutralization. Am J pathol 1998, 153(4):1249-1256.
• [7]Huang S, Robinson JB, DeGuzman A, Bucana CD, Fidler IJ: Blockade of nuclear factor-κB signaling inhibits angiogenesis and tumorigenicity of human ovarian cancer cells by suppressing expression of vascular endothelial growth factor and interleukin 8. Cancer Res 2000, 60(19):5334-5339.
• [8]Goodenough DA, Goliger JA, Paul DL: Connexins, connexons, and intercellular communication. Annu Rev Biochem 1996, 65:475-502.
• [9]Vinken M, de Kock J, Oliveira AG, Menezes GB, Cogliati B, Dagli ML, Vanhaecke T, Rogiers V: Modifications in connexin expression in liver development and cancer. Cell Commun Adhes 2012, 19(3–4):55-62.
• [10]Sun Y, Zhao X, Yao Y, Qi X, Yuan Y, Hu Y: Connexin 43 interacts with Bax to regulate apoptosis of pancreatic cancer through a gap junction-independent pathway. Int J Oncol 2012, 41(3):941-948.
• [11]Li Z, Zhou Z, Welch DR, Donahue HJ: Expressing connexin 43 in breast cancer cells reduces their metastasis to lungs. Clin Exp Metastasis 2008, 25(8):893-901.
• [12]Zhang YW, Kaneda M, Morita I: The gap junction-independent tumor-suppressing effect of connexin 43. J Biol Chem 2003, 278(45):44852-44856.
• [13]Corteggio A, Florio J, Roperto F, Borzacchiello G: Expression of gap junction protein connexin 43 in bovine urinary bladder tumours. J Comp Pathol 2011, 144(1):86-90.
• [14]Umhauer S, Ruch RJ, Fanning J: Gap junctional intercellular communication and connexin 43 expression in ovarian carcinoma. Am J Obstet Gynecol 2000, 182(5):999-1000.
• [15]Spinella F, Rosano L, Di Castro V, Nicotra MR, Natali PG, Bagnato A: Endothelin-1 decreases gap junctional intercellular communication by inducing phosphorylation of connexin 43 in human ovarian carcinoma cells. J Biol Chem 2003, 278(42):41294-41301.
• [16]Jou YS, Matesic D, Dupont E, Lu SC, Rupp HL, Madhukar BV, Oh SY, Trosko JE, Chang CC: Restoration of gap-junctional intercellular communication in a communication-deficient rat liver cell mutant by transfection with connexin 43 cDNA. Mol Carcinog 1993, 8(4):234-244.
• [17]Pimentel RC, Yamada KA, Kléber AG, Saffitz JE: Autocrine regulation of myocyte Cx43 expression by VEGF. Circ Res 2002, 90(6):671-677.
• [18]Iyer RK, Odedra D, Chiu LL, Vunjak-Novakovic G, Radisic M: Vascular endothelial growth factor secretion by nonmyocytes modulates Connexin-43 levels in cardiac organoids. Tissue Eng Part A 2012, 18(17–18):1771-1783.
• [19]Jiang J, Slivova V, Harvey K, Valachovicova T, Sliva D: Ganoderma lucidum suppresses growth of breast cancer cells through the inhibition of Akt/NF-kappaB signaling. Nutr Cancer 2004, 49(2):209-216.
• [20]Loganathan J, Jiang J, Smith A, Jedinak A, Thyagarajan-Sahu A, Sandusky GE, Nakshatri H, Sliva D: The mushroom Ganoderma lucidum suppresses breast-to-lung cancer metastasis through the inhibition of pro-invasive genes. Int J Oncol 2014, 44(6):2009-2015.
• [21]Thyagarajan-Sahu A, Lane B, Sliva D: ReishiMax, mushroom based dietary supplement, inhibits adipocyte differentiation, stimulates glucose uptake and activates AMPK. BMC Complement Altern Med. 2011, 11:74. BioMed Central Full Text
• [22]Wang HH, Kung CI, Tseng YY, Lin YC, Chen CH, Tsai CH, Yeh HI: Activation of endothelial cells to pathological status by down-regulation of connexin43. Cardiovasc Res 2008, 79(3):509-518.
• [23]Peterson SM, Freeman JL: RNA isolation from embryonic zebrafish and cDNA synthesis for gene expression analysis. J Vis Exp 2009, 30:1470.
• [24]Jayson GC, Kohn EC, Kitchener HC, Ledermann JA: Ovarian cancer. Lancet 2014, 384(9951):1376-1388.
• [25]Kim HW, Won KS, Zeon SK, Ahn BC, Gayed IW: Peritoneal carcinomatosis in patients with ovarian cancer: enhanced CT versus 18 F-FDG PET/CT. Clin Nucl Med 2013, 38(2):93-97.
• [26]Zytoon AA, Murakami K, Eid H, El-Gammal M: High impact of FDG-PET/CT in diagnostic strategies for ovarian cancer. Acta Radiol 2013, 54(3):340-348.
• [27]Klostergaard J, Parga K, Raptis RG: Current and future applications of magnetic resonance imaging (MRI) to breast and ovarian cancer patient management. P R Health Sci J 2010, 29(3):223-231.
• [28]Diamandis EP: Mass spectrometry as a diagnostic and a cancer biomarker discovery tool opportunities and potential limitations. Mol Cell Proteomics 2004, 3(4):367-378.
• [29]Tekpli X, Rivedal E, Gorria M, Landvik NE, Rissel M, Dimanche-Boitrel MT, Baffet G, Holme JA, Lagadic-Gossmann D: The B[a]P-increased intercellular communication via translocation of connexin-43 into gap junctions reduces apoptosis. Toxicol Appl Pharmacol 2010, 242(2):231-240.
• [30]Li L, Wang L, Zhang W, Tang B, Zhang J, Song H, Yao D, Tang Y, Chen X, Yang Z, Wang G, Li X, Zhao J, Ding H, Reed E, Li QQ: Correlation of serum VEGF levels with clinical stage, therapy efficacy, tumor metastasis and patient survival in ovarian cancer. Anticancer Res 2004, 24(3b):1973-1979.