可利霉素通过调控巨噬细胞极化影响黑色素瘤的增殖<sup>*</sup>

doi:10.13523/j.cb.2203018

中国生物工程杂志

2022, Vol. 42

Issue (7): 12-23 DOI: 10.13523/j.cb.2203018

研究报告

可利霉素通过调控巨噬细胞极化影响黑色素瘤的增殖^*

杨子荣¹,杨璇¹,倪婷婷²,潘聪³,谭诗生^1,^2,^**(

),王姿^1,^2,^**(

)

1. 贵州大学医学院贵阳 550025
2. 贵州省人民医院肿瘤科贵阳 550002
3. 四川大学生物治疗国家重点实验室成都 610041

Carrimycin Affects Melanoma Proliferation by Regulating Macrophage Polarization

Zi-rong YANG¹,Xuan YANG¹,Ting-ting NI²,Cong PAN³,Shi-sheng TAN^1,^2,^**(

),Zi WANG^1,^2,^**(

)

1. Medicine College, Guizhou University, Guiyang 550025, China
2. Department of Oncology, Guizhou Provincial People’s Hospital, Guiyang 550002, China
3. State Key Laboratory of Biotherapy, Sichuan University, Chengdu 610041, China

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摘要： 目的探讨可利霉素(CAM)是否通过调节巨噬细胞极化,继而影响黑色素瘤的发生发展。方法通过实时荧光定量聚合酶链反应(Q-RT-PCR)和Western blot检测CAM对巨噬细胞极化的影响;流式细胞术和CCK-8法检测CAM对小鼠体内外巨噬细胞吞噬和增殖的影响;构建B16-F10细胞系荷瘤模型评价CAM对黑色素瘤的抗肿瘤作用。结果 CAM可上调M1巨噬细胞TNF-α和iNOS的水平,下调M2中Arg-1的水平;上调p-STAT1表达,下调p-STAT3表达。CAM抑制黑色素瘤的发生发展,抑瘤率为41.6%,促进M1巨噬细胞数量增加 (P<0.05)。结论 CAM促进体内M1巨噬细胞数量增多,抑制黑色素瘤的进展,提示CAM可能通过诱导巨噬细胞向M1巨噬细胞极化发挥抗肿瘤作用。

关键词： 可利霉素; 黑色素瘤; 抗肿瘤; 巨噬细胞; 极化

Abstract:

To investigate whether carrimycin (CAM) affects the occurrence and development of melanoma by regulating the polarization of macrophages, and the following related cell biology assays were used to examine its function. Methods: The effect of CAM on macrophage polarization was detected by real-time quantitative polymerase chain reaction (Q-RT-PCR) and Western blot. Flow cytometry and Cell Counting Kit-8 were used to detect the effect of CAM on mouse macrophages in vitro and in vivo phagocytosis and proliferation. Cell line-derived xenograft model was constructed via B16-F10 to evaluate the anti-tumor effect of CAM on melanoma. Results: In the mRNA level, CAM could up-regulate the levels of TNF-α and iNOS in M1 and down-regulate the level of Arg-1 in M2. In the protein level, CAM can increase the expression of p-STAT1 and decrease the expression of p-STAT3. In the cell line-derived xenograft model, these data shown that the occurrence and CAM development of melanoma was inhibited after CAM treatment, the tumor inhibition rate was 41.6%, and promoted the increase of the number of M1 macrophages (P<0.05). Conclusion: CAM promotes the increase in the number of M1 macrophages in vivo and inhibits the progression of melanoma, suggesting that CAM may achieve anti-tumor effects by inducing the polarization of macrophages to M1.

Key words: Carrimycin Melanoma Anti-tumor Macrophage Polarization

收稿日期: 2022-03-08 出版日期: 2022-08-03

ZTFLH:

Q813

基金资助: *贵州省海外高层次人才创新创业基金([2019] 03);国家自然科学基金(81860535);贵州省自然科学基金(黔科合基础[2020]1Y339(黔科合基础[2020]1Y339)

通讯作者: 谭诗生,王姿 E-mail: wangzi@gz5055.com;tssh18018@126.com

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引用本文:

杨子荣,杨璇,倪婷婷,潘聪,谭诗生,王姿. 可利霉素通过调控巨噬细胞极化影响黑色素瘤的增殖^*[J]. 中国生物工程杂志, 2022, 42(7): 12-23.

Zi-rong YANG,Xuan YANG,Ting-ting NI,Cong PAN,Shi-sheng TAN,Zi WANG. Carrimycin Affects Melanoma Proliferation by Regulating Macrophage Polarization. China Biotechnology, 2022, 42(7): 12-23.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.2203018 或 https://manu60.magtech.com.cn/biotech/CN/Y2022/V42/I7/12

表1 引物序列

图1 CAM 对体外巨噬细胞吞噬和增殖的影响

图2 定量-RT PCR 分析 CAM 对 RAW246.7 细胞极化的影响

图3 定量-RT PCR 分析 CAM 对原代巨噬细胞极化的影响

图4 Western blot检测CAM对巨噬细胞极化通路蛋白的作用

图5 CAM对体内单核细胞和巨噬细胞增殖的影响

图6 CAM 在小鼠黑色素瘤模型中的抗肿瘤作用

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