Gamma linolenic acid suppresses hypoxia-induced gastric cancer cell growth and epithelial-mesenchymal transition by inhibiting the Wnt/b-catenin signaling pathway
Abstract
Introduction. Gastric cancer is one of the most common malignancies in China and the fifth most common cancer in the world. Gamma linolenic acid (GLA) was reported to have anti-inflammatory and anti-cancer effects. The purpose of this research was to investigate the effect and mechanism of GLA on gastric cancer cell growth under hypoxic conditions.
Material and methods. The hypoxia models of SGC-7901 and MGC-803 cells were established, and then were exposed to different concentrations of 50, 100 or 200 μM GLA. MTT assay, colony formation assay, wound healing assay and transwell assay were used to investigate the effects of GLA treatment on gastric cancer cell growth under hypoxia (1% O2). The expression of apoptosis- and epithelial-mesenchymal transition (EMT)-related proteins was detected by qPCR and western blot.
Results. GLA treatment significantly decreased viability and inhibited colony formation (p < 0.05, p < 0.01) of SGC-7901 and MGC-803 cells under hypoxia. Western blotting analysis showed that GLA treatment decreased the expression of proliferating cell nuclear antigen (PCNA), microchromosome maintenance complex component 2 (MCM-2) and anti-apoptotic protein Bcl-2, while increased the expression of pro-apoptotic proteins (Bax and Cleaved Caspase-3) (p < 0.05 and p < 0.01). In addition, Wound healing analysis and Transwell assays showed that GLA treatment inhibited the migration and invasion of SGC-7901 and MGC-803 cells in a dose-dependent manner (p < 0.01). Western blotting analysis showed that GLA treatment increased the expression of epithelial marker proteins (g-catenin and E-cadherin), while decreased the expression of stromal and extracellular matrix marker proteins (fibronectin, Snail and b-catenin) (p < 0.01). Further analyses showed that GLA treatment decreased the expression of b-catenin in Wnt/b-catenin pathway (p < 0.01). Moreover, exogenous Wnt3a reversed the inhibitory effect of GLA on b-catenin expression, and further reversed the inhibitory effect of GLA on gastric cancer cell growth and EMT markers (p < 0.05, p < 0.01).
Conclusion. These findings suggest that GLA should be tested in animal models and in clinical studies as a potentially effective bioactive phytochemical substance for the treatment of gastric cancer.
Keywords: Gamma linolenic acidgastric cancerSGC-7901 cellsMGC-803 cellshypoxiacell growthWnt/b-cateninEMTapoptosis
References
- Wu C, Li M, Meng H, et al. Analysis of status and countermeasures of cancer incidence and mortality in China. Sci China Life Sci. 2019; 62(5): 640–647.
- Ferlay J, Soerjomataram I, Dikshit R, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 2015; 136(5): E359–E386.
- Ferlay J, Shin HR, Bray F, et al. Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer. 2010; 127(12): 2893–2917.
- Ohtsu A. Chemotherapy for metastatic gastric cancer: past, present, and future. J Gastroenterol. 2008; 43(4): 256–264.
- Liu Na, Wang X. [Current status and research progress of perioperative chemotherapy in advanced gastric cancer]. Zhonghua Wei Chang Wai Ke Za Zhi. 2015; 18(10): 983–985.
- Allemani C, Weir HK, Carreira H, et al. CONCORD Working Group. Global surveillance of cancer survival 1995-2009: analysis of individual data for 25,676,887 patients from 279 population-based registries in 67 countries (CONCORD-2). Lancet. 2015; 385(9972): 977–1010.
- Patrad E, Niapour A, Farassati F, et al. Combination treatment of all-trans retinoic acid (ATRA) and γ-secretase inhibitor (DAPT) cause growth inhibition and apoptosis induction in the human gastric cancer cell line. Cytotechnology. 2018; 70(2): 865–877.
- Makuuchi R, Sugisawa N, Kaji S, et al. Enhanced recovery after surgery for gastric cancer and an assessment of preoperative carbohydrate loading. Eur J Surg Oncol. 2017; 43(1): 210–217.
- Zhao J, Wang G, Jiang ZW, et al. Patients Administered Neoadjuvant Chemotherapy Could be Enrolled into an Enhanced Recovery after Surgery Program for Locally Advanced Gastric Cancer. Chin Med J (Engl). 2018; 131(4): 413–419.
- Tegels JJW, De Maat MFG, Hulsewé KWE, et al. Improving the outcomes in gastric cancer surgery. World J Gastroenterol. 2014; 20(38): 13692–13704.
- Shimizu N, Hatao F, Fukatsu K, et al. Results of a nationwide questionnaire-based survey on nutrition management following gastric cancer resection in Japan. Surg Today. 2017; 47(12): 1460–1468.
- Klek S, Scislo L, Walewska E, et al. Enriched enteral nutrition may improve short-term survival in stage IV gastric cancer patients: A randomized, controlled trial. Nutrition. 2017; 36: 46–53.
- Tasset-Cuevas I, Fernández-Bedmar Z, Lozano-Baena MD, et al. Protective effect of borage seed oil and gamma linolenic acid on DNA: in vivo and in vitro studies. PLoS One. 2013; 8(2): e56986.
- Guil-Guerrero JL, Garcı́a-Maroto F, Campra-Madrid P, et al. Occurrence and characterization of oils rich in γ-linolenic acid Part II: fatty acids and squalene from Macaronesian Echium leaves. Phytochemistry. 2000; 54(5): 525–529.
- Bégin ME, Ells G, Das UN, et al. Differential killing of human carcinoma cells supplemented with n-3 and n-6 polyunsaturated fatty acids. J Natl Cancer Inst. 1986; 77(5): 1053–1062.
- Bégin ME. Effects of polyunsaturated fatty acids and of their oxidation products on cell survival. Chem Phys Lipids. 1987; 45(2-4): 269–313.
- Dai J, Shen J, Pan W, et al. Effects of polyunsaturated fatty acids on the growth of gastric cancer cells in vitro. Lipids Health Dis. 2013; 12: 71.
- Menendez JA, Colomer R, Lupu R. Omega-6 polyunsaturated fatty acid gamma-linolenic acid (18:3n-6) is a selective estrogen-response modulator in human breast cancer cells: gamma-linolenic acid antagonizes estrogen receptor-dependent transcriptional activity, transcriptionally represses estrogen receptor expression and synergistically enhances tamoxifen and ICI 182,780 (Faslodex) efficacy in human breast cancer cells. Int J Cancer. 2004; 109(6): 949–954.
- Menéndez JA, Ropero S, del Barbacid MM, et al. Synergistic interaction between vinorelbine and gamma-linolenic acid in breast cancer cells. Breast Cancer Res Treat. 2002; 72(3): 203–219.
- Robbins M, Ali K, McCaw R, et al. gamma-linolenic acid (GLA)-mediated cytotoxicity in human prostate cancer cells. Adv Exp Med Biol. 1999; 469: 499–504.
- Wang X, Lin H, Gu Y. Multiple roles of dihomo-γ-linolenic acid against proliferation diseases. Lipids Health Dis. 2012; 11: 25.
- Cui H, Han F, Zhang L, et al. Gamma linolenic acid regulates PHD2 mediated hypoxia and mitochondrial apoptosis in DEN induced hepatocellular carcinoma. Drug Des Devel Ther. 2018; 12: 4241–4252.
- Cui H, Han F, Zhang L, et al. Gamma linolenic acid regulates PHD2 mediated hypoxia and mitochondrial apoptosis in DEN induced hepatocellular carcinoma. Drug Des Devel Ther. 2018; 12: 4241–4252.
- Chas M, Goupille C, Arbion F, et al. Low eicosapentaenoic acid and gamma-linolenic acid levels in breast adipose tissue are associated with inflammatory breast cancer. Breast. 2019; 45: 113–117.
- Zhao S, Min P, Liu L, et al. NEDD9 Facilitates Hypoxia-Induced Gastric Cancer Cell Migration via MICAL1 Related Rac1 Activation. Front Pharmacol. 2019; 10: 291.
- Zhang Qi, Bai X, Chen W, et al. Wnt/β-catenin signaling enhances hypoxia-induced epithelial-mesenchymal transition in hepatocellular carcinoma via crosstalk with hif-1α signaling. Carcinogenesis. 2013; 34(5): 962–973.
- Xu Yi, Qi J, Yang X, et al. Free radical derivatives formed from cyclooxygenase-catalyzed dihomo-γ-linolenic acid peroxidation can attenuate colon cancer cell growth and enhance 5-fluorouracil's cytotoxicity. Redox Biol. 2014; 2: 610–618.
- Yang X, Xu Yi, Gao Di, et al. Dihomo-γ-linolenic acid inhibits growth of xenograft tumors in mice bearing human pancreatic cancer cells (BxPC-3) transfected with delta-5-desaturase shRNA. Redox Biol. 2019; 20: 236–246.
- Roy S, Rawat AK, Sammi SR, et al. Alpha-linolenic acid stabilizes HIF-1 α and downregulates FASN to promote mitochondrial apoptosis for mammary gland chemoprevention. Oncotarget. 2017; 8(41): 70049–70071.
- Rani A, Roy S, Singh M, et al. α-Chymotrypsin regulates free fatty acids and UCHL-1 to ameliorate N-methyl nitrosourea induced mammary gland carcinoma in albino wistar rats. Inflammopharmacology. 2016; 24(5): 277–286.
- Balakumar P, Taneja G. Fish oil and vascular endothelial protection: bench to bedside. Free Radic Biol Med. 2012; 53(2): 271–279.
- Saha B, Patro BS, Koli M, et al. -4,4'-Dihydroxystilbene (DHS) inhibits human neuroblastoma tumor growth and induces mitochondrial and lysosomal damages in neuroblastoma cell lines. Oncotarget. 2017; 8(43): 73905–73924.
- Roy S, Singh M, Rawat A, et al. GLA supplementation regulates PHD2 mediated hypoxia and mitochondrial apoptosis in DMBA induced mammary gland carcinoma. Int J Biochem Cell Biol. 2018; 96: 51–62.
- Murakami K, Wu Y, Imaizumi T, et al. DEC1 promotes hypoxia-induced epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells. Biomed Res. 2017; 38(4): 221–227.
- Lv WL, Liu Q, An JH, et al. Scutellarin inhibits hypoxia-induced epithelial-mesenchymal transition in bladder cancer cells. J Cell Physiol. 2019; 234(12): 23169–23175.
- Deng M, Xue YJ, Xu LR, et al. Chrysophanol Suppresses Hypoxia-Induced Epithelial-Mesenchymal Transition in Colorectal Cancer Cells. Anat Rec (Hoboken). 2019; 302(9): 1561–1570.
- Ruan J, Zhang L, Yan L, et al. Inhibition of hypoxia-induced epithelial mesenchymal transition by luteolin in non-small cell lung cancer cells. Mol Med Rep. 2012; 6(1): 232–238.
- Jo H, Lee J, Jeon J, et al. The critical role of glucose deprivation in epithelial-mesenchymal transition in hepatocellular carcinoma under hypoxia. Sci Rep. 2020; 10(1): 1538.
- Tam SY, Wu VWC, Law HKW. JNK Pathway Mediates Low Oxygen Level Induced Epithelial-Mesenchymal Transition and Stemness Maintenance in Colorectal Cancer Cells. Cancers (Basel). 2020; 12(1).
- Zhu X, Lu X. MiR-423-5p inhibition alleviates cardiomyocyte apoptosis and mitochondrial dysfunction caused by hypoxia/reoxygenation through activation of the wnt/β-catenin signaling pathway via targeting MYBL2. J Cell Physiol. 2019; 234(12): 22034–22043.
- Qian YY, Shi YY, Lu SH, et al. Extracts of Celastrus Orbiculatus Inhibit Cancer Metastasis by Down-regulating Epithelial-Mesenchymal Transition in Hypoxia-Induced Human Hepatocellular Carcinoma Cells. Chin J Integr Med. 2019; 25(5): 334–341.
- Su J, Su Bo, Xia H, et al. RORα Suppresses Epithelial-to-Mesenchymal Transition and Invasion in Human Gastric Cancer Cells via the Wnt/β-Catenin Pathway. Front Oncol. 2019; 9: 1344.
- Wang Xi, Lu B, Dai C, et al. Caveolin-1 Promotes Chemoresistance of Gastric Cancer Cells to Cisplatin by Activating WNT/β-Catenin Pathway. Front Oncol. 2020; 10: 46.
- Zeng S, Seifert AM, Zhang JQ, et al. Wnt/β-catenin Signaling Contributes to Tumor Malignancy and Is Targetable in Gastrointestinal Stromal Tumor. Mol Cancer Ther. 2017; 16(9): 1954–1966.
- Kim M, Kim SH, Lim JW, et al. Lycopene induces apoptosis by inhibiting nuclear translocation of β-catenin in gastric cancer cells. J Physiol Pharmacol. 2019; 70(4).
- Yuan H, Zhang J, Li F, et al. Sinomenine exerts antitumour effect in gastric cancer cells via enhancement of miR-204 expression. Basic Clin Pharmacol Toxicol. 2019; 125(5): 450–459.
- Hou G, Yuan X, Li Yi, et al. Cardamonin, a natural chalcone, reduces 5-fluorouracil resistance of gastric cancer cells through targeting Wnt/β-catenin signal pathway. Invest New Drugs. 2020; 38(2): 329–339.
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