| 孙艳华,李明,徐建立,等.紫草素通过激活自噬相关的 E1连接酶 7信号通路诱导结肠癌细胞自噬[J].安徽医药,2020,24(12):2336-2342. |
| 紫草素通过激活自噬相关的 E1连接酶 7信号通路诱导结肠癌细胞自噬 |
| Shikonin induces autophagy in colon cancer cells by activating the autophagy?related E1 ligase 7 signaling pathway |
| |
| DOI：10.3969/j.issn.1009?6469.2020.12.002 |
| 中文关键词: 结肠肿瘤 紫草素 细胞自噬 泛素蛋白连接酶类 细胞凋亡 细胞增殖 程序性细胞死亡受体 1 |
| 英文关键词: Colonic neoplasms Shikonin Autophagy Programmed cell death 1 receptor |
| 基金项目:2019年度河北省医学科学研究重点课题计划(20191249) |
|
| 摘要点击次数: 2241 |
| 全文下载次数: 722 |
| 中文摘要: |
| 目的研究紫草素对人结肠癌细胞系 SNU?407增殖和自噬的影响，及其潜在的分子机制。方法通过细胞计数试剂盒 ? 8(CCK?8)检测细胞存活率，用 Hoechst 33258染色法和流式细胞术分析细胞生长情况，采用蛋白质印迹法(Western Blot)分析不同紫草素浓度(0μmol/L、10 μmol/L、20 μmol/L和 30 μmol/L)处理前后自噬相关的 E1连接酶 7(ATG7)、丝裂原活化蛋白激酶(MAPK)和死亡受体(Caspase?8，Caspase?3，PARP)蛋白表达情况。结果紫草素以时间和剂量依赖性方式诱导 SNU?407细胞凋亡和自噬，在紫草素处理的 SNU?407细胞中观察到 ATG7表达水平显著升高；与 0 μmol/L的紫草素相比， 10 μmol/L、20 μmol/L和 30 μmol/L紫草素处理后 SNU?407细胞的 ATG7/自噬相关的 E1连接酶 5(ATG5)相对表达量分别显著增加［(176±9)%、(212±13)%和(293±11)%］。与 GFPi(阴性对照质粒 pSUPER_GFP siRNA)阴性对照组相比， ATG被沉默的 shATG7(ATG7的 RNA干扰质粒 pSUPER_ATG7 siRNA)组 LC3?Ⅱ/LC3?Ⅰ比值显著低于 GFPi阴性对照组［(8.25±1.36)比(97.45±1.98)，P＜0.001］ATG7沉默后抑制了紫草素诱导的 SNU?407细胞自噬；荧光显微镜结果和流式细胞术分析证实紫草素诱导结肠癌细胞凋亡；质印迹法印迹分析显示紫草素通过 MAPK活化诱导结肠癌细胞凋亡，激活死亡受体(FADD)调节细胞凋亡蛋白(Caspase?8，Caspase?3，PARP)表达。此外，紫草素通过增加 ATG7的表达水平，进而下调促进细胞自噬的 p62表达。结论紫蛋白，草素通过激活 ATG7信号通路诱导人结肠癌细胞系 SNU?407细胞凋亡和自噬。 |
| 英文摘要: |
| Objective To elucidate the effects of shikonin on proliferation and autophagy of human colon cancer cell line SNU?407，and to explore its molecular mechanism of potential action.Methods Cell Counting Kit?8(CCK?8)was used to detect cell viability，Hoechst 33258 staining and flow cytometry were used to analyze cell growth，and Western Blot was used to analyze different shikonin concentrations(0 μmol/L，10 μmol/L，20 μmol/L and 30 μmol/L)autophagy?related E1 ligase 7(ATG7)，mito? gen?activated protein kinase(MAPK) and death receptor(Caspase?8，Caspase?3，PARP) protein expression before and after treatment Happening.Results The results showed that shikonin induced apoptosis of SNU?407 cells in a time?and dose?dependent manner，and ATG7 expression level was significantly increased in shikonin?treated SNU?407 cells.Compared with 0 μmol/L shiko? nin，the relative expression of ATG7/autophagy?related E1 ligase 5(ATG5)in SNU?407 cells was significantly increased by(176± 9)%，(212±13)% and(293±11)% after treatment with 10 μmol/L，20 μmol/L and 30 μmol/L shikonin.Compared with GFPi nega? tive control group(the negative control plasmid pSUPER_GFP siRNA)，the LC3?Ⅱ/LC3?Ⅰ ratio of the shATG7 group(ATG7 RNA interference plasmid pSUPER_ATG7 siRNA)was significantly lower than that of the GFPi negative control group［(8.25± 1.36)vs.(97.45±1.98)P＜0.001］，silencing of ANU7 inhibited shikonin?induced apoptosis of SNU?407 cells.Furtherly，fluores? cencemicroscopyresultsa，nd flow cytometry analysis confirmed that shikonin induced apoptosis in colorectal cancer cells.In addi?tion，Western blotting analysis showed that shikonin induced apoptosis of colon cancer cells through MAPK activation，and activat? ed death receptor(FADD)to regulate the expression of apoptosis proteins(Caspase?8，Caspase?3，PARP).In addition，shikonin in? creases the expression level of ATG7，thereby down?regulating the expression of p62 that promotes autophagy.Conclusion Based on the above results，we suggest that shikonin promotes the initiation of autophagosomes by activating the ATG7 signaling pathway to induce apoptosis and autophagy in human colon cancer cell SNU?407. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
