3-甲基腺嘌呤对人耐药大肠癌SW480细胞增殖及自噬活性的影响

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (841 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的：研究自噬抑制剂3-甲基腺嘌呤（3-MA）对耐5-氟尿嘧啶（5-Fu）结肠癌SW480细胞增殖及自噬活性的影响，探讨自噬抑制剂在逆转结直肠癌耐药中的作用。方法：浓度递增筛选法诱导耐5-Fu（100μg/L）的SW480细胞株，用10μmol/L浓度的3-MA处理后，再用5-Fu（浓度200μg/L）作用24 h。采用CCK-8法检测细胞增殖情况，流式细胞仪检测细胞凋亡率，丹酰戊二胺（MDC）染色和电镜检测细胞自噬活性变化。结果：3-MA对耐药细胞自噬活性的抑制率达89.7%，增殖抑制率为9.6%，凋亡率为5.7%，与对照组比较差异具有统计学意义（P＜0.05）。经3-MA处理后，5-Fu对耐药SW480细胞增殖抑制率达71.6%±2.9%，凋亡率达36.6%±2.9%，而单用5-Fu细胞增殖抑制率仅为23.6%±2.9%，凋亡率仅为10.3%±2.5%，3-MA预处理后5-Fu对耐药细胞的增殖抑制率及凋亡率均明显增强（P＜0.05），自噬泡数量显著减少，自噬活性明显降低（P＜0.05）。结论：3-MA单用时仅有较弱的抗肿瘤作用，与5-FU联用时耐药细胞的增殖及自噬活性被显著抑制，增强了SW480耐药细胞对5-Fu的敏感性，提高了化疗效果。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	胡万乐
	王玲琴
	陈苏苹
	陈荣

关键词 ：结直肠肿瘤, 自噬, 3-甲基腺嘌呤, 氟尿嘧啶, 耐药

Abstract：Objective: To investigate the effects of 3-methyladenine on chemotherapy sensitivity of 5-Furesistant colon cancer cell line SW480 and reversing the 5-Furesistant in colon cancer. Methods: Stepwise 5-Fu selection was used to establish the 5-Fu resistant colon cancer cells (100μg/L). The cells were dealed with 3-methyladenine (10μmol/L) fistly and 5-Fu (200μg/L) for 24 hours. Cell proliferation inhibition rate was measured with CCK-8. Apoptosis rate was detected with flow cytometer. Autophagic vacuoles in cells were observed by means of transmission electron microscopy and monodansylcadaverine (MDC). Results: The inhibition rate of 3-methyladenine in the 5-Fu resistant cell autophagy activity was 89.7%, cell proliferation was 9.6%, but apoptosis rate was just 5.7%, were obviously higher than that in 5-Fu resistant cells (P<0.05). After dealing with 3-methyladenine(10μmol/L) fistly and 5-Fu (200μg/L) for 24 hours, the level of cell proliferation inhibition rate was 71.6%±2.9% and the apoptosis rate was 36.6%±2.9%, while dealing with 5-Fu (200μg/L) for 24 hours, the level of cell proliferation inhibition rate was 23.6%±2.9% and the apoptosis rate was 10.3%±2.5%. After dealing with 3-methyladenine, the cell proliferation inhibition rate and apoptosis rate were markedly higher than that of 5-Fu only. Conclusion: The anticancer capacity of 3-methyladenine in colon cancer is weak, but it can reverse the 5-Fu-resistant in colon cancer partly, 5-Fu-induced apoptosis in 5-Fu resistant colon cancer cells may be enhanced by the inhibitor of autophagy.

Key words： colonrectal neoplasms autophagy 3-methyladenine fluorouracil drug resistance

收稿日期: 2013-10-25

基金资助:温州市科技计划基金资助项目（Y20120012）

通讯作者: 陈荣，主任医师，副教授，Email：cr13806887833@ 163.com。

作者简介: 胡万乐（1978-），女，浙江温州人，主治医师

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2014/V44/I3/189

[1] Shi Y, Tang B, Yu PW, et al. Autophagy protects against oxaliplatin-induced cell death via ER stress and ROS in Caco-2 cells[J]. PLoS One, 2012, 7(11): 51076.

[2] Fu J, Liu ZG, Liu XM, et al. Glioblastoma stem cells resistant to temozolomide-induced autophagy[J]. Chin Med J (Engl),2009, 122(11): 1255-1259.

[3] Cui J, Gong Z, Shen HM. The role of autophagy in liver cancer: molecular mechanisms and potential therapeutic targets[J]. Biochim Biophys Acta, 2013, 1836(1): 15-26.

[4] 汪鑫, 刘天助, 郭洪波, 等. 恶性胶质瘤耐药细胞通过Beclin 1上调自噬抵抗替莫唑胺化疗[J]. 中华神经医学杂志, 2013, 12 (8): 757-762.

[5] 苑博, 康劲松, 杨晓春, 等. 自噬在紫杉醇诱导人卵巢癌顺铂耐药细胞死亡中的作用[J]. 中国实验诊断学, 2013, 17(1):46-48.

[6] 胡万乐, 何占红, 蒋芳, 等. 5-Fu诱导人大肠癌SW480细胞株耐药与自噬活性的变化[J]. 中国中西医结合外科杂志,2013, 19(3): 279-284.

[7] Hirosako K, Imasato H, Hirota Y,et al. 3-Methyladenine specifically inhibits retrograde transport of cation-independent mannose 6-phosphate/insulin-like growth factor II receptor from the early endosome to the TGN[J]. Biochem Biophys Res Commun, 2004, 316(3): 845-852.

[8] Mizushima N, Klionsky DJ. Protein turnover via autophagy:implications for metabolism[J]. Annu Rev Nutr, 2007, 27:19-40.

[9] Loewith R, Jacinto E, Wullschleger S, et al. Two TOR complexes, only one of which is rapamycin sensitive, have distinct roles in cell growth control[J]. Mol Cell, 2002, 10 (3): 457-468.

[10] Groves B, Abrahamsen H, Clingan H, et al. Aninhibitory role of the G-protein regulator AGS3 in mTOR-dependent macroautophagy[J]. PLoS One, 2010, 26, 5(1): e8877.

[11] Broadley K, Larsen L, Herst PM, et al. The novel phloroglucinol PMT7 kills glycolytic cancer cells by blocking autophagy and sensitizing to nutrient stress[J]. J Cell Biochem,2011, 112(7): 1869-1879.

[12] Ahn JH, Lee M. Suppression of autophagy sensitizes multidrug resistant cells towards Src tyrosine kinase specific inhibitor PP2[J]. Cancer Lett, 2011, 310(2): 188-197.

[13] 龙璐璐, 许文林, 沈慧玲, 等. YB-1抑制三氧化二砷诱导的胃癌细胞自噬机制[J]. 山东医药, 2012, 52(13): 4-6.