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陈娟,徐琳本,蒋益兰,翦林宏,曾宏亮,谭小宁.健脾消癌方调控巨噬细胞M2极化对结直肠癌细胞侵袭作用的研究[J].湖南中医药大学学报,2022,42(12):2002-2007[点击复制] |
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| 健脾消癌方调控巨噬细胞M2极化对结直肠癌细胞侵袭作用的研究 |
| 陈娟,徐琳本,蒋益兰,翦林宏,曾宏亮,谭小宁 |
| (湖南省中医药研究院, 湖南 长沙 410006) |
| 摘要: |
| 目的 探讨健脾消癌方调控巨噬细胞M2极化对结直肠癌HCT116细胞侵袭转移能力的影响。方法 使用佛波酯诱导THP-1细胞成M0型巨噬细胞,通过Transwell法构建HCT116细胞与M0型巨噬细胞共培养体系,设立空白血清对照组、健脾消癌方含药血清组、IL-4+空白血清组、IL-4+健脾消癌方含药血清组。RT-PCR法检测巨噬细胞M2极化相关基因C-C基序趋化因子22(C-C motif chemokine22, CCL22)、精氨酸酶-1(arginase-1, Arg-1)表达,Transwell法检测HCT116侵袭转移能力,Western blot法检测HCT116中E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)及基质金属蛋白酶9(matrix metalloproteinase, MMP-9)蛋白表达水平。结果 与空白血清对照组比较,IL-4+空白血清组能上调CCL22、Arg-1表达(P<0.01),下调HCT116细胞E-cadherin表达、上调Vimentin、MMP-9表达(P<0.01),增加结直肠癌HCT116细胞侵袭数(P<0.01);与空白血清对照组比较,健脾消癌方含药血清组能下调CCL22、Arg-1表达(P<0.05),上调HCT116细胞E-cadherin表达(P<0.05),下调Vimentin表达(P<0.05),减少结直肠癌HCT116细胞侵袭数(P<0.01);与IL-4+空白血清组比较,IL-4+健脾消癌方含药血清组可下调CCL22(P<0.05)、Arg-1(P<0.01)表达,上调HCT116细胞E-cadherin表达(P<0.05),下调Vimentin、MMP-9表达(P<0.05),减少结直肠癌HCT116细胞侵袭数(P<0.01)。结论 健脾消癌方能一定程度上抑制巨噬细胞M2极化,阻断HCT116细胞上皮间质转化进程而抑制结直肠癌细胞侵袭转移。 |
| 关键词: 健脾消癌方 结直肠癌 巨噬细胞 M2极化 共培养 侵袭转移 上皮间质转化 |
| DOI：10.3969/j.issn.1674-070X.2022.12.008 |
| 投稿时间：2022-07-23 |
| 基金项目:国家自然科学基金项目(81774287); 湖南省自然科学基金项目(2020JJ4413); 湖南省教育厅科学研究项目(19C1412) |
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| Effects of Jianpi Xiao'ai Formula on the invasion of colorectal cancer cells by regulating M2 polarization of macrophages |
| CHEN Juan,XU Linben,JIANG Yilan,JIAN Linhong,ZENG Hongliang,TAN Xiaoning |
| (Hunan Academy of Chinese Medicine, Changsha, Hunan 410006, China) |
| Abstract: |
| Objective To investigate the effects of Jianpi Xiao'ai Formula on the invasion ability of HCT116 colorectal cancer cells by regulating macrophage M2 polarization.Methods THP-1 cells were induced to M0 macrophages by phorbol ester. Then, HCT116 cells and M0 type macrophages were co-cultured by Transwell. The co-cultured cells were randomly divided into blank serum control group, Jianpi Xiao'ai Formula serum group, IL-4 + blank serum group, and IL-4 + Jianpi Xiao'ai Formula serum group. The RT-PCR was used to detect the expression of C-C motif chemokine22(CCL22) and arginase-1(Arg-1), the related genes of M2 polarization in macrophages, and Transwell was applied to detect the invasion and metastasis of HCT116. The expression levels of E-cadherin, Vimentin and matrix metalloproteinase(MMP-9) in HCT116 were determined by Western blot.Results Compared with blank serum control group, IL-4 + blank serum group could up-regulate the expression of CCL22 and Arg-1(P <0.01), down-r egulate the expression of E-cadherin and up-regulate the expression of Vimentin and MMP-9 in HCT116 cells(P<0.01), and increased the invasion number of colorectal cancer HCT116 cells(P<0.01). Compared with blank serum control group, Jianpi Xiao'ai Formula serum group showed the lower expression of CCL22 and Arg-1(P<0.05), higher E-cadherin expression in HCT116 cells(P<0.05), lower Vimentin expression(P<0.05), and less colorectal cancer HCT116 cell invasion(P<0.01). Compared IL-4 + blank serum group, IL-4 + Jianpi Xiao'ai Formula serum group could down-regulate CCL22(P<0.05), and Arg-1 expression(P<0.01), up-regulated E-cadherin expression in HCT116 cells(P<0.05), down-regulated the expression of Vimentin and MMP-9(P<0.05), and decreased the number of colorectal cancer HCT116 cell invasion(P<0.01).Conclusion Jianpi Xiao'ai Formula could inhibit M2 polarization of macrophages, block EMT process of HCT116 cells and inhibit the invasion and metastasis of colorectal cancer cells. |
| Key words: Jianpi Xiao'ai Formula colorectal cancer macrophage M2 polarization co-culture invasion and metastasis epithelial-mesenchymal transition |
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