大蒜素对小鼠黑色素瘤B16F10细胞体外增殖和迁移能力的影响

doi:10.11843/j.issn.0366-6964.2021.03.027

Abstract

Abstract: The study aimed to investigate the effects of different concentrations of allicin on the expression of microphthalmia-associated transcription factor (MITF) and its effects on the proliferation and migration of B16F10 cells, which provide a new idea for the treatment of animal melanoma in pet and animal husbandry. B16F10 cells were divided into control group (0 μg·mL^-1) and experimental group (5, 10, 15, 20 μg·mL^-1) supplemented with different concentrations of allicin. The expression of MITF mRNA in B16F10 cells was detected by qRT-PCR and the expression of MITF protein was detected by Western blot. The cell growth rate was calculated by using the absorbance of cells at 515 nm, and then the inhibition rate of different concentrations of allicin on cell proliferation was determined. Wound healing was used to evaluate the effects of different concentrations of allicin on the migration of B16F10 cells after 42 hours. The results showed that the expression of MITF mRNA decreased gradually with the increase of allicin concentration, while Western blot results showed that the expression of MITF protein decreased in a concentration-dependent manner. Allicin at the concentrations of 5, 10, 15, 20 μg·mL^-1 could significantly inhibit the proliferation of B16F10 cells (P<0.001), and the inhibition rate increased with the increase of allicin concentration. Wound healing results showed that allicin significantly inhibited the cell migration at the concentration of 10 μg·mL^-1, 15 μg·mL^-1 and 20 μg·mL^-1. This experiment showed that allicin could affect the expression of the melanoma-specific marker gene MITF and inhibit the proliferation and migration of B16F10 cells.

Key words: B16F10, MITF, allicin, proliferation, migration

CLC Number:

S857.4

ZHU Zhiwei, LAN Wutao, MA Yueyue, LI Pengfei, LI Hong, ZHANG Lei, SI Wenrui, FENG Jianghao. Effects of Allicin on Proliferation and Migration of Mouse Melanoma B16F10 Cells in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 831-838.

References 29

[1]	董小龙. 皮肤恶性黑色素瘤诊断及外科治疗策略[J]. 中国美容医学, 2016, 25(6):108-112.DONG X L. Progress in the diagnosis and surgical treatments for cutaneous malignant melanoma[J]. Chinese Journal of Aesthetic Medicine, 2016, 25(6):108-112. (in Chinese)
[2]	秦毓敏, 钟友刚. 犬黑色素瘤研究进展[C]//第六届西部宠物医师大会论文集. 成都:中国畜牧兽医学会, 2017:4.QIN Y M, ZHONG Y G. Advances in canine melanoma research[C]//The 6th Western Small Animal Veterinarian Congress Collected Papers. Chengdu:China Animal Husbandry and Veterinary Association, 2017:4. (in Chinese)
[3]	BLANKENSTEIN S A, VAN AKKOOI A C J. Adjuvant systemic therapy in high-risk melanoma[J]. Melanoma Res, 2019, 29(4):358-364.
[4]	刘文艳, 李园, 蔡永强, 等. 黑色素细胞中MITF相关信号通路调控因子的研究进展[J]. 畜牧与兽医, 2017, 49(8):125-128.LIU W Y, LI Y, CAI Y Q, et al. Research progress of the regulators involved in MITF-related signaling pathways in melanocytes[J]. Animal Husbandry & Veterinary Medicine, 2017, 49(8):125-128. (in Chinese)
[5]	ULLRICH N, LÖFFEK S, HORN S, et al. MITF is a critical regulator of the carcinoembryonic antigen-related cell adhesion molecule 1(CEACAM1) in malignant melanoma[J]. Pigment Cell Melanoma Res, 2015, 28(6):736-740.
[6]	SIEGEL R L, MILLER K D, JEMAL A. Cancer statistics, 2016[J]. CA Cancer J Clin, 2016, 66(1):7-30.
[7]	赵美娟, 户晶晶, 倪辉, 等. 黑色素生成信号通路研究进展[J]. 生物工程学报, 2019, 35(9):1633-1642.ZHAO M J, HU J J, NI H, et al. Research progress in melanogenesis signaling pathway[J]. Chinese Journal of Biotechnology, 2019, 35(9):1633-1642. (in Chinese)
[8]	韩涛, 陈世军, 喻昌盛, 等. 马常见皮肤肿瘤的诊断与治疗[J]. 现代农业科技, 2014(15):298-299.HAN T, CHEN S J, YU C S, et al. Diagnosis and treatment of equine common skin tumors[J]. Modern Agricultural Science and Technology, 2014(15):298-299. (in Chinese)
[9]	宋桂太, 宋维臣, 庄欠海, 等. 手术切除牛黑色素瘤1例[J]. 山东畜牧兽医, 1998(5):42-43.SONG G T, SONG W C, ZHUANG Q H, et al. Surgical resection of bovine melanoma in 1 case[J]. Shandong Journal of Animal Science and Veterinary Medicine, 1998(5):42-43. (in Chinese)
[10]	RAMOS-VARA J A, BEISSENHERZ M E, MILLER M A, et al. Retrospective study of 338 canine oral melanomas with clinical, histologic, and immunohistochemical review of 129 cases[J]. Vet Pathol, 2000, 37(6):597-608.
[11]	韩庆功, 刘超, 陈金山, 等. 猪恶性黑色素瘤的病理学观察[J]. 中国兽医杂志, 2015, 51(8):37-38.HAN Q G, LIU C, CHEN J S, et al. Pathological observation of malignant melanoma in pigs[J]. Chinese Journal of Veterinary Medicine, 2015, 51(8):37-38. (in Chinese)
[12]	蔡彤, 邓佳佳, 彭西. 一例犬口腔恶性黑色素瘤的病理诊断及治疗[J]. 畜禽业, 2019, 30(8):119.CAI T, DENG J J, PENG X. Pathological diagnosis and treatment of a case of oral malignant melanoma in dogs[J]. Livestock and Poultry Industry, 2019, 30(8):119. (in Chinese)
[13]	张琦, 白喜云, 段步婷, 等. 一例原发性猫眼黑色素瘤的组织病理学分析[C]//中国畜牧兽医学会兽医病理学分会第二十五次学术交流会、中国病理生理学会动物病理学专业委员会第二十四次学术研讨会、中国实验动物学会实验病理学专业委员会第四次学术研讨会、中国兽医病理学家第四次研讨会论文集. 天津:中国畜牧兽医学会兽医病理学分会, 2019:107.ZHANG Q, BAI X Y, DUAN B T, et al. Histopathological analysis of a case of primary cat eye melanoma[C]//The 25th Academic Exchange Meeting of Veterinary Pathology Branch of Chinese Animal Husbandry and Veterinary Association, the 24th Academic Seminar of Animal Pathology Professional Committee of Chinese Society of Pathophysiology, the Fourth Academic Seminar of Experimental Pathology Professional Committee of Chinese Society of Experimental Animals, and the Fourth Symposium of Chinese Veterinary Pathologist. Tianjin:Veterinary Pathology Branch of Chinese Animal Husbandry and Veterinary Association, 2019:107. (in Chinese)
[14]	LV J P, FU Y, CAO Y, et al. Isoliquiritigenin inhibits melanogenesis, melanocyte dendricity and melanosome transport by regulating ERK-mediated MITF degradation[J]. Exp Dermatol, 2020, 29(2):149-157.
[15]	杨鹏欣, 刘蕾, 马来记, 等. 黑素合成相关MITF基因及其表达研究概况[J]. 日用化学工业, 2017, 47(1):46-51.YANG P X, LIU L, MA L J, et al. Situation of research work with respect to melanogenesis-associated MITF gene and its expression[J]. China Surfactant Detergent & Cosmetics, 2017, 47(1):46-51. (in Chinese)
[16]	YUN C Y, ROH E, KIM S H, et al. Stem cell factor-inducible MITF-M expression in therapeutics for acquired skin hyperpigmentation[J]. Theranostics, 2020, 10(1):340-352.
[17]	余翔. 白化荣昌猪黑色素生成通路相关基因变异分析[D]. 雅安:四川农业大学, 2011.YU X. Analysis of gene variation related to melanin production pathway in Rongchang albino pig[D]. Yaan:Sichuan Agricultural University, 2011. (in Chinese)
[18]	杨玉静, 张丹瑾, 聂瑞强, 等. 绵羊MITF-M在黑素细胞中过表达后的功能分析[J]. 中国农业科学, 2016, 49(21):4214-4221.YANG Y J, ZHANG D J, NIE R Q, et al. The function analysis of over-expression of oar MITF-M in melanocytes[J]. Scientia Agricultura Sinica, 2016, 49(21):4214-4221. (in Chinese)
[19]	VLČKOVÁ K, VACHTENHEIM J, RÉDA J, et al. Inducibly decreased MITF levels do not affect proliferation and phenotype switching but reduce differentiation of melanoma cells[J]. J Cell Mol Med, 2018, 22(4):2240-2251.
[20]	PHUNG B, SUN J M, SCHEPSKY A, et al. Abstract 3093:elucidation of c-KIT- dependent signaling to MITF[J]. Cancer Res, 2012, 72(8 Suppl):3093.
[21]	FRANZONI A, MARKOVA-CAR E, DEVIĆ-PAVLIĆ S, et al. A polymorphic GGC repeat in the NPAS2 gene and its association with melanoma[J]. Exp Biol Med (Maywood), 2017, 242(15):1553-1558.
[22]	TIFFEN J, WILSON S, GALLAGHER S J, et al. Somatic copy number amplification and hyperactivating somatic mutations of EZH2 correlate with DNA methylation and drive epigenetic silencing of genes involved in tumor suppression and immune responses in melanoma[J]. Neoplasia, 2016, 18(2):121-132.
[23]	姬凯元, 范瑞文, 张俊珍, 等. 小鼠正常黑色素细胞与黑色素瘤细胞(B16)mRNA表达差异分析[J]. 中国生物化学与分子生物学报, 2019, 35(1):92-100.JI K Y, FAN R W, ZHANG J Z, et al. Differential expression of mRNA between murine melanocytes and B16 melanoma cells[J]. Chinese Journal of Biochemistry and Molecular Biology, 2019, 35(1):92-100. (in Chinese)
[24]	吴达军, 许峰. 晚期恶性黑色素瘤的新药治疗进展[J]. 华西医学, 2016, 31(12):2079-2083.WU D J, XU F. Advances in new drug therapy for advanced malignant melanoma[J]. West China Medical Journal, 2016, 31(12):2079-2083. (in Chinese)
[25]	SALEHI B, ZUCCA P, ORHAN I E, et al. Allicin and health:a comprehensive review[J]. Trends Food Sci Technol, 2019, 86:502-516.
[26]	林思祥, 杨桂青, 王园园, 等. 黑蒜提取液对恶性黑色素瘤B16细胞增殖与凋亡影响及其机制的研究[J]. 中华中医药杂志, 2014, 29(4):1240-1243.LIN S X, YANG G Q, WANG Y Y, et al. Study on the mechanism and effect of aged black garlic extract on proliferation and apoptosis of malignant melanoma B16 cells[J]. China Journal of Traditional Chinese Medicine and Pharmacy, 2014, 29(4):1240-1243. (in Chinese)
[27]	李建波, 樊汉利, 王建祥. 大蒜素激活Wnt/β-连环蛋白通路抑制A549肺癌小鼠转移侵袭的研究[J]. 中华实验外科杂志, 2019, 36(11):2058-2060.LI J B, FAN H L, WANG J X. Allicin activates Wingless/β-catenin pathway to inhibit metastasis and invasion of A549 lung cancer mice[J]. Chinese Journal of Experimental Surgery, 2019, 36(11):2058-2060. (in Chinese)
[28]	吴淑芬, 李志晋. 大蒜素抑制内质网自噬影响胃癌SGC-7901细胞增殖和凋亡的研究[J]. 肿瘤药学, 2019, 9(2):226-230.WU S F, LI Z J. Effect of allicin on the proliferation and apoptosis of gastric cancer cells SGC-7901 through inhibiting autophagy of endoplasmic reticulum[J]. Anti-tumor Pharmacy, 2019, 9(2):226-230. (in Chinese)
[29]	SINNAMON A J, NEUWIRTH M G, YALAMANCHI P, et al. Association between patient age and lymph node positivity in thin melanoma[J]. JAMA Dermatol, 2017, 153(9):866-873.

Effects of Allicin on Proliferation and Migration of Mouse Melanoma B16F10 Cells in vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 29

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	MIAO Shu, AN Jishan, WANG Zuo, XIAO Dingfu, LAN Xinyi, LIU Lei, SHEN Weijun, WAN Fachun. Leucine Promotes the Proliferation of Bovine Myoblasts through PI3K-AKT Signaling Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 142-152.
[2]	DUAN Xiangru, KANG Jia, YANG Ruochen, SHAN Xinyu, LI Taichun, ZHAO Wen, ZHANG Yingjie, LIU Yueqin. Effect of L-cysteine on Proliferation, Apoptosis and the Secretion of Steroid Hormone in Ovine Ovarian Granulosa Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 179-191.
[3]	YUAN Heling, FANG Ci, HUANG Jinhu, WANG Xiaoming, XIAO Wenjun, LIU Ruiting, SHI Rongmei, WANG Liping. Effects of Allicin on mRNA Expression of CYP1A2 and Its Enzyme Kinetics in Broilers Liver Tissue [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3905-3915.
[4]	ZHANG Wanfeng, ZHAO Tianzhi, LI Jiao, YOU Ziwei, YANG Yang, CAI Chunbo, GAO Pengfei, CAO Guoqing, GUO Xiaohong, LI Bugao. Study on NR2F2 Gene Regulating Proliferation and Apoptosis of Porcine PK15 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3242-3251.
[5]	HE Mingyang, MA Yujing, WANG Yong, YANG Ruochen, LIU Yueqin, ZHANG Yingjie, DUAN Chunhui. Effects of Melatonin on Proliferation, Apoptosis of Ovarian Granulosa Cells, and Its Secretion of Steroid Hormones of Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3313-3324.
[6]	SONG Meijun, HAO Kexing, HAI Siyu, CHEN Yan, WANG Jing, HU Guangdong. Effects of SRIF-14 on Proliferation and Apoptosis of Endometrial Epithelial Cells in Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3325-3334.
[7]	WANG Wanjie, CHEN Nanzhu, ZOU Huiying, ZHOU Xinyi, HAO Haisheng, PANG Yunwei, ZHU Huabin, ZHAO Xueming, YU Dawei, DU Weihua. Effects of Histone Methyltransferase ASH1L Overexpression on Proliferation and Apoptosis of Bovine Cumulus Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3358-3368.
[8]	ZHANG Peng, WANG Mingxiu, JING Kemin, PENG Wei, TIAN Yuan, LI Yuqian, FU Changqi, SHU Shi, ZHONG Jincheng, CAI Xin. Abnormal Expression of FGFs/FGFRs and Their Mediated Signaling Pathway Genes Affect the Proliferative Activity of Undifferentiated Spermatogonia in Cattleyak [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2886-2897.
[9]	XU Tiantian, ZHANG Tongtong, WANG Meng, WANG Xin. Transcription Factor Foxq1 Affects the Proliferation of Hair Follicle Stem Cells in Cashmere Goats via WNT/β-catenin Signaling Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2653-2661.
[10]	YUN Jiale, LIU Chang, HUANG Xiaoyu, LIU Qiaoxia, SHI Mingyue, LI Wenxia, NIU Jin, WANG Shouyuan, GAO Pengfei, GUO Xiaohong, LI Bugao, LU Chang, CAO Guoqing. miR-145-5p Inhibits the Proliferation and Differentiation of Porcine Skeletal Muscle Satellite Cells by Targeting IGF1R-Mediated AKT Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1893-1904.
[11]	WANG Kaiming, YU Zonggang, XU Xueli, AI Nini, LI Xintong, HE Jun, TAO Deng, ZHANG Shuo, MA Haiming, ZHANG Yuebo. Effect of Interfering Mtmr3 on C2C12 Cells Proliferation and Differentiation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1478-1489.
[12]	REN Xiaoli, FAN Yuying, HUANGFU Heping, LIU Yun, SHI Dongmei. Effect of GSK126 on Epithelial-mesenchymal Transition of Canine Mammary Tumor Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1721-1729.
[13]	YANG Guang, XU Jing, LI Xin, HU Debao, GUO Yiwen, DING Xiangbin, GUO Hong, ZHANG Linlin. Effect of Interfering lncbMD on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1015-1025.
[14]	YANG Xiaowei, ZHAO Ziliang, FU Yu, YU Zixiao, ZHAO Yongju. Effects of TET1 Gene on the Proliferation of Mouse uNK Cells and the Transcriptional Level of IFN-γ, VEGF-C and TGF-β1 [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1221-1228.
[15]	SUN Wenye, LI Jianji, WANG Heng, DONG Junsheng, LI Jun, CUI Luying. Effect of Met-enkephalin on Lipopolysaccharide-stimulated Proliferation of Bovine Endometrial Stromal Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1229-1239.