| 杨颖颖,邱炜.乳酸诱导巨噬细胞 M2型极化对黑色素瘤细胞增殖、凋亡、迁移和侵袭的影响[J].安徽医药,2023,27(11):2145-2149. |
| 乳酸诱导巨噬细胞 M2型极化对黑色素瘤细胞增殖、凋亡、迁移和侵袭的影响 |
| Effects of lactic acid-induced M2-type polarization of macrophages on proliferation, apoptosis, migration and invasion of melanoma cells |
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| DOI：10.3969/j.issn.1009-6469.2023.11.006 |
| 中文关键词: 乳酸 巨噬细胞 M2型极化 黑色素瘤 增殖 凋亡 迁移 侵袭 |
| 英文关键词: Lactic acid Macrophage M2-type polarization Melanoma Proliferation Apoptosis Migration Invasion |
| 基金项目:国家自然科学基金资助项目( 31960206) |
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| 中文摘要: |
| 目的探究乳酸( LA)是否可通过调控巨噬细胞 M2型极化影响黑色素瘤细胞增殖、凋亡、迁移和侵袭。方法 2021年 1月至 2022年 2月采用细胞计数试剂盒( CCK-8)法检测乳酸对细胞的毒性；使用乳酸干预巨噬细胞后的细胞上清液培养黑色素瘤细胞，经 CCK-8法和流式细胞术检测细胞增殖和凋亡；经划痕愈合实验和 Transwell法检测细胞迁移和侵袭。通过光学显微镜对乳酸干预后的巨噬细胞的形态进行观察，并通过 ELISA检测巨噬细胞极化相关因子的表达情况。实时荧光定量逆转录聚合酶链式反应( qRT-PCR)检测乳酸对巨噬细胞中肿瘤蛋白翻译调节因子 1(Tpt1)、肺腺癌转移相关转录本 1(Malat1)表达的影响。结果乳酸对巨噬细胞 Raw264.7和黑色素瘤细胞 B16F10均无明显毒性( P>0.05)。 5、10、20 mmol/L乳酸干预 Raw264.7细胞后的上清液后， B16F10细胞活力［(125.36±7.88)%、(144.36±8.65)%、(167.28±10.12)%比( 100.00±0.00)%］划痕愈合率［( 39.21±3.15)%、(52.65±3.87)%、(64.24±6.12)%比( 25.26±2.23)%］、侵袭细胞数［( 89.36±8.12)个、(113.05±10.03)个、(131.25±11.14)个比( 45.36±4.21)个］增高，凋亡率［(41.23±4.02)%、(29.26±3.14)%、(18.64±2.09)%比( 57.26±4.25)%］下降( P<0.05)。5、10、20 mmol/L乳酸作用 Raw264.7细胞后，细胞形态出现 M2型形态变化， M1型相关分子表达差异无统计学意义( P>0.05)，M2型相关因子血管内皮生长因子( VEGF)、白细胞介素 -10(IL-10)［( 47.26±4.02)ng/L、(65.32±5.48)ng/L、(79.36±7.12) ng/L比( 27.25±2.21)ng/L］、 CD163表达均显著增多( P<0.05)。乳酸对 Raw264.7细胞中 Tpt1、Malat1表达具有明显的促进作用(P<0.05)。结论乳酸诱导巨噬细胞 M2型极化促进黑色素瘤恶性生物学发展，且 Tpt1、Malat1表达可能参与此过程。 |
| 英文摘要: |
| Objective To investigate whether lactic acid (LA) can affect melanoma cell proliferation, apoptosis, migration and invasion by regulating macrophage M2-type polarization.Methods The toxicity of LA to cells was detected by CCK-8 method from January 2021 to February 2022;the melanoma cells were cultured with the cell supernatant after LA intervening macrophages, and cell proliferation and apoptosis were detected by CCK-8 method and flow cytometry; the cell migration and invasion were detected by scratchhealing assay and Transwell assay. The morphology of macrophages after LA intervention was observed by light microscope, and the expression of macrophage polarization-related factors was detected by ELISA. The effects of LA on the expressions of tumor protein translation regulator 1 (Tpt1) and lung adenocarcinoma metastasis-associated transcript 1 (Malat1) in macrophages were measured by qRT-PCR. Results LA had no obvious toxicity to macrophage Raw264.7 and melanoma cell B16F10(P>0.05). After the supernatant ofRaw264.7 cells was treated with 5,10 and 20 mmol/L LA,B16F10 cell activity [(125.36±7.88)%,(144.36±8.65)%,(167.28±10.12)% vs. (100.00±0.00)% ], scratch healing rate [(39.21±3.15)% , (52.65±3.87)% , (64.24±6.12)% vs. (25.26±2.23)% ] and invasion cell number [(89.36±8.12) individual, (113.05±10.03) individual, (131.25±11.14) individual vs. (45.36±4.21) individual] increased, the apoptosis rate [(41.23±4.02)% , (29.26±3.14)% , (18.64±2.09)% vs. (57.26±4.25)% ] decreased(P<0.05). After treating Raw264.7 cells with 5, 10 and 20 mmol/L LA,the cell morphology showed M2-type morphological changes,and there was no significant difference in the expression of M1-type-related molecules (P>0.05),but the expressions of M2-type-related factors vascular endothelial growth factor (VEGF),interleukin-10 (IL-10) [(47.26±4.02) ng/L,(65.32±5.48) ng/L,(79.36±7.12) ng/L vs. (27.25±2.21) ng/L] and CD163 were greatly increased(P<0.05).LA greatly promoted the expressions of Tpt1 and Malat1 in Raw264.7 cells (P<0.05).Conclusion LA induces M2-type polarization of macrophages to promote the malignant biological development of melanoma, and the expressions of Tpt1 and Malat1 may beinvolved in this process. |
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