喹乙醇诱导HepG2 细胞自噬的实验研究

doi:10.3969/j.issn.1004-616x.2013.03.002

Abstract

Abstract:

OBJECTIVE: To investigate the effect of olaquindox on autophagy in human hepatoma G2 (HepG2) cells. METHODS：The HepG2 cells were treated with different concentrations (0，200，400，800 μg/mL) of olaquindox for 24 h. Then autophagy was analyzed by fluorescence microscope and flow cytometer following staining with monodansylcadaverine (MDC). HepG2 cells were treated with different concentrations (0，200，400，800 μg/mL) of olaquindox for 24 h or with 400 μg/mL olaquindox for different periods (0，1，3，6，12，24 h). Then the expressions of LC3-Ⅱ and Beclin 1 were determined by Western blot. RESULTS：Olaquindox-treated cells exhibited higher fluorescent density and more MDC-labeled particles in HepG2 cells compared with the control group. The percentage of MDC-positive cells was obviously increased after treatment with different concentrations (200，400，800 μg/mL) of olaquindox for 24 h. (P<0.05 or P<0.01). The expressions of LC3-Ⅱ and Beclin 1 increased with different concentrations and different time points of olaquindox treatment. Moreover，compared with the control group，the expression of LC3-Ⅱ and Beclin 1 were increased significantly with treatment of 400 μg/mL olaquindox for 24 h (P<0.01). CONCLUSION： Olaquindox enhanced the autophagic process in HepG2 cells.

Key words: olaquindox, autophagy, MDC, LC3-Ⅱ, Beclin 1

ZHAO Dong-xu，ZHANG Chao-ming，TANG Shu-sheng， WANG Cong-cong，ZHANG Shen，XIAO Xi-long. Autophagic process induced by olaquindox in HepG2 cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.03.002.

References

[1] 王树槐. 喹乙醇诱发CHL细胞染色体畸变试验[J]. 中国兽药杂志，1993，27(4)：27-29.

[2] Mantovani A. The role of multigeneration studies in safety assessment of residues of veterinary drugs and additives[J]. Annali dell'Istituto Superiore di Sanita，1992，28(4)：429-435.

[3] Yoshimura H，Nakamura M，Koeda T，et al. Mutagenicities of carbadox and olaquindox-growth promoters for pigs[J]. Mutat Res，1981，90 (1)：49-55.

[4] Zou JJ，Chen Q，Jin X，et al. Olaquindox induces apoptosis through the mitochondrial pathway in HepG2 cells[J]. Toxicology，2011，285 (3)：104-113.

[5] 赵文霞，张朝明，汤树生，等. p38MAPK在喹乙醇诱导HepG2 细胞凋亡中的作用[J]. 癌变?畸变?突变，2012， 24 (1)：10-13.

[6] Mizushima N，Ohsumi Y，Yoshimori T. Autophagosome formation in mammalian cells[J]. Cell Struct Funct，2002，27 (6)：421-429.

[7] Lum JJ，Bauer DE，Kong M，et al. Growth factor regulation of autophagy and cell survival in the absence of apoptosis[J]. Cell，2005，120 (2)：237-248.

[8] Karantza-Wadsworth V，Patel S，Kravchuk O，et al. Autophagy mitigates metabolic stress and genome damage in mammary tumorigenesis[J]. Gene Dev，2007，21 (13)：1621-1635.

[9] Sakaki K，Wu J，Kaufman RJ. Protein kinase C theta is required for autophagy in response to stress in the endoplasmic reticulum [J]. J Biol Chem，2008，283 (22)：15370-15380.

[10] 尤寿江，石际俊，张艳林，等. ROS介导的自噬及其在相关疾病中的作用[J]. 中国病理生理杂志，2011，27 (1)：187-190.

[11] Guo WJ，Ye SS，Cao N，et al. ROS-mediated autophagy was involved in cancer cell death induced by novel copper(II) complex[J]. Experiment Toxicol Pathol，2010， 62 (5)：577-582.

[12] Bolt AM，Klimecki WT. Autophagy in toxicology：self-consumption in times of stress and plenty[J]. J Appl Toxicol，2012，32 (7)： 465-479.

[13] 汪开毓，耿毅，叶仕根，等. 鲤慢性喹乙醇中毒的病理学和组织残留[J]. 水产学报，2003，27 (1)：75-82.

[14] 尹荣焕，白文林，吴长德，等. 喹乙醇对小鼠肝脏和肾脏的毒性作用研究[J]. 畜牧兽医学报，2008，39 (7)：974-979.

[15] Klionsky DJ，Abdalla FC，Abeliovich H. Guidelines for the use and interpretation of assays for monitoring autophagy[J]. Autophagy，2012，8 (4)：1-100.

[16] Biederbick A，Kern HF，Elsasser HP. Monodansylcadaverine (MDC) is a specific in-vivo marker for autophagic vacuoles[J]. Eur J Cell Biol，1995，66 (1)：3-14.

[17] Mizushima N，Yoshimori T. How to interpret LC3 immunoblotting [J]. Autophagy，2007，3 (6)：542-545.

[18] Yue ZY，Jin SK，Yang CW，et al. Beclin 1，an autophagy gene essential for early embryonic development，is a haploin-sufficient tumor suppressor[J]. Proc Natl Acad Sci USA，2003， 100 (25)：15077-15082.

[19] Wirawan E，Vande Walle L，Kersse K，et al. Caspase-mediated cleavage of Beclin-1 inactivates Beclin-1-induced autophagy and enhances apoptosis by promoting the release of proapoptotic factors from mitochondria[J]. Cell Death Dis，2010，1 (1)：1-10.

[20] Li J，Hou N，Faried A，et al. Inhibition of autophagy by 3-MA Enhances the effect of 5-FU-induced apoptosis in colon cancer cells[J]. Ann Surg Oncol，2009，16 (3)：761-771.

[21] Liu WT，Lin CH，Hsiao M，et al. Minocycline inhibits the growth of glioma by inducing autophagy[J]. Autophagy，2011， 7 (2)：166-175.

[22] Camougrand N，Grelaud-Coq A，Marza E，et al. The product of the UTH1 gene，required for Bax-induced cell death in yeast，is involved in the response to rapamycin[J]. Mol Microbiol，2003，47 (2)：495-506.

[23] 金昕，袁栋栋，陈秀玲，等. 磷酸腺苷激活的蛋白激酶(AMPK)参与了曲格列酮诱导的HeLa细胞自噬和凋亡[J]. 微生物学报，2012，(7)：840-849.

[24] Eskelinen EL. The dual role of autophagy in cancer[J]. Curr Opin Pharmacol，2011，11 (4)：294-300.

[25] Chen YQ，Klionsky DJ. The regulation of autophagy - unanswered questions[J]. J Cell Sci，2011，124 (2)：161-170.

Autophagic process induced by olaquindox in HepG2 cells

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