[目的]探讨miR-346靶向基质金属蛋白酶-9(MMP-9)对绒毛膜癌细胞株JAR细胞侵袭增殖的影响。[方法]设绒毛膜癌细胞株JAR组、miR-NC组、miR-346 mimics组、miR-346 inhibitor组,培养72 h。MTT法检测绒毛膜癌细胞株JAR活力、癌细胞细胞集落...[目的]探讨miR-346靶向基质金属蛋白酶-9(MMP-9)对绒毛膜癌细胞株JAR细胞侵袭增殖的影响。[方法]设绒毛膜癌细胞株JAR组、miR-NC组、miR-346 mimics组、miR-346 inhibitor组,培养72 h。MTT法检测绒毛膜癌细胞株JAR活力、癌细胞细胞集落,流式细胞仪检测细胞凋亡水平,Transwell实验测定细胞侵袭水平,RT-PCR法及蛋白电泳法测定miR-346、MMP-9水平。[结果]与绒毛膜癌细胞株JAR组和miR-NC组比较,miR-346 mimics组OD值、存活率、细胞集落、穿膜数、迁移距离、MMP-9 mRNA和蛋白水平降低[0.73±0.03、0.72±0.03 vs 0.41±0.04,(78.24±6.25、76.37±7.35 vs 42.74±7.34)%,619.47±89.35、614.88±75.27 vs 239.32±35.53,693.85±184.74、702.28±111.52 vs 293.85±95.28,(100.25±18.55、98.54±17.45 vs 68.25±12.24)μm, 3.85±0.25、3.90±0.23 vs 2.04±0.24,0.90±0.22、0.89±0.23 vs 0.37±0.26],细胞凋亡率、miR-346水平[(12.38±1.37、12.26±1.20 vs 25.39±1.47)%,3.17±0.33、3.15±0.36 vs 5.68±0.37]升高(P<0.05),miR-346 inhibitor组OD值、存活率、细胞集落、穿膜数、迁移距离、MMP-9 mRNA和蛋白水平(0.90±0.04,86.95%±6.24%,1062.28±102.79,1917.34±425.35,198.25μm±25.66μm, 4.63±0.28,1.48±0.34)升高,细胞凋亡率、miR-346水平(6.62±0.54,1.42±0.38)降低(P<0.05)。[结论] miR-346对绒毛膜癌细胞株JAR细胞侵袭、增殖具有明显抑制作用,其机制可能与miR-346靶向抑制MMP-9表达有关。展开更多
[目的]探讨乳腺癌细胞分泌琥珀酸盐促进巨噬细胞极化的潜在机制。[方法]不同乳腺癌细胞系或正常乳腺上皮细胞的培养上清培养人巨噬细胞后,分析TAM标志物Arg1、Fizz1、Mgl1的表达以及巨噬细胞极化为TAM水平。检测乳腺癌细胞和正常乳腺上...[目的]探讨乳腺癌细胞分泌琥珀酸盐促进巨噬细胞极化的潜在机制。[方法]不同乳腺癌细胞系或正常乳腺上皮细胞的培养上清培养人巨噬细胞后,分析TAM标志物Arg1、Fizz1、Mgl1的表达以及巨噬细胞极化为TAM水平。检测乳腺癌细胞和正常乳腺上皮细胞中琥珀酸盐的水平并评估其对巨噬细胞极化为TAM的影响。分子对接和表面等离子共振分析琥珀酸与琥珀酸受体(SUCNR1)的结合位点。[结果]乳腺癌细胞上清培养的巨噬细胞中TAM标志物的表达水平上升(1.00±0.08 vs 19.71±1.80,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.10 vs 4.89±0.44,P<0.05)。乳腺癌细胞上清中琥珀酸的水平显著上升[(0.09±0.01 vs 0.63±0.04)mmol/L,P<0.05]。不同浓度琥珀酸处理后,巨噬细胞中TAM标志物的表达水平呈浓度依赖性上升(1.00±0.01 vs 26.14±0.21,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.04 vs 4.10±0.23,P<0.05)分子对接和表面等离子共振显示琥珀酸盐与SUCNR1结合于R81、N173和S178位点。[结论]乳腺癌细胞分泌的琥珀酸盐与巨噬细胞SUCNR1的R81、N173和S178结合后促进了巨噬细胞发生M2极化为TAM(1.00±0.10 vs 4.89±0.44)。展开更多
The JAR choriocarcinoma cell line share many of the characteristics of early placental trophoblast cells including the invasion properties. Matrix metallo-proteinases (MMPs), the main actors of matrix proteolysis, are...The JAR choriocarcinoma cell line share many of the characteristics of early placental trophoblast cells including the invasion properties. Matrix metallo-proteinases (MMPs), the main actors of matrix proteolysis, are involved in normal invasion as well as in the invasive character of tumor cells and the metastase formation. Tumor necrosis factor-α (TNF-α) is present in the placental environment and TNF-α levels are elevated in some placental pathologies. In the present work, we addressed whether TNF-α is a modulator of JAR cell MMP secretion. Following TNF-α stimulation, zymographic analysis showed the increased secretion of the active form of MMP-2 and to a lesser extent proMMP-2 and MMP-9. In addition, MMP-2 gene expression only increased slightly whereas MMP-9 and TIMP-1 transcripts were undetectable. This suggests that TNF-α may modulate the secretion of MMPs independently of MMP gene expression control.展开更多
文摘[目的]探讨miR-346靶向基质金属蛋白酶-9(MMP-9)对绒毛膜癌细胞株JAR细胞侵袭增殖的影响。[方法]设绒毛膜癌细胞株JAR组、miR-NC组、miR-346 mimics组、miR-346 inhibitor组,培养72 h。MTT法检测绒毛膜癌细胞株JAR活力、癌细胞细胞集落,流式细胞仪检测细胞凋亡水平,Transwell实验测定细胞侵袭水平,RT-PCR法及蛋白电泳法测定miR-346、MMP-9水平。[结果]与绒毛膜癌细胞株JAR组和miR-NC组比较,miR-346 mimics组OD值、存活率、细胞集落、穿膜数、迁移距离、MMP-9 mRNA和蛋白水平降低[0.73±0.03、0.72±0.03 vs 0.41±0.04,(78.24±6.25、76.37±7.35 vs 42.74±7.34)%,619.47±89.35、614.88±75.27 vs 239.32±35.53,693.85±184.74、702.28±111.52 vs 293.85±95.28,(100.25±18.55、98.54±17.45 vs 68.25±12.24)μm, 3.85±0.25、3.90±0.23 vs 2.04±0.24,0.90±0.22、0.89±0.23 vs 0.37±0.26],细胞凋亡率、miR-346水平[(12.38±1.37、12.26±1.20 vs 25.39±1.47)%,3.17±0.33、3.15±0.36 vs 5.68±0.37]升高(P<0.05),miR-346 inhibitor组OD值、存活率、细胞集落、穿膜数、迁移距离、MMP-9 mRNA和蛋白水平(0.90±0.04,86.95%±6.24%,1062.28±102.79,1917.34±425.35,198.25μm±25.66μm, 4.63±0.28,1.48±0.34)升高,细胞凋亡率、miR-346水平(6.62±0.54,1.42±0.38)降低(P<0.05)。[结论] miR-346对绒毛膜癌细胞株JAR细胞侵袭、增殖具有明显抑制作用,其机制可能与miR-346靶向抑制MMP-9表达有关。
文摘[目的]探讨乳腺癌细胞分泌琥珀酸盐促进巨噬细胞极化的潜在机制。[方法]不同乳腺癌细胞系或正常乳腺上皮细胞的培养上清培养人巨噬细胞后,分析TAM标志物Arg1、Fizz1、Mgl1的表达以及巨噬细胞极化为TAM水平。检测乳腺癌细胞和正常乳腺上皮细胞中琥珀酸盐的水平并评估其对巨噬细胞极化为TAM的影响。分子对接和表面等离子共振分析琥珀酸与琥珀酸受体(SUCNR1)的结合位点。[结果]乳腺癌细胞上清培养的巨噬细胞中TAM标志物的表达水平上升(1.00±0.08 vs 19.71±1.80,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.10 vs 4.89±0.44,P<0.05)。乳腺癌细胞上清中琥珀酸的水平显著上升[(0.09±0.01 vs 0.63±0.04)mmol/L,P<0.05]。不同浓度琥珀酸处理后,巨噬细胞中TAM标志物的表达水平呈浓度依赖性上升(1.00±0.01 vs 26.14±0.21,P<0.05),并且巨噬细胞极化为TAM的水平上升(1.00±0.04 vs 4.10±0.23,P<0.05)分子对接和表面等离子共振显示琥珀酸盐与SUCNR1结合于R81、N173和S178位点。[结论]乳腺癌细胞分泌的琥珀酸盐与巨噬细胞SUCNR1的R81、N173和S178结合后促进了巨噬细胞发生M2极化为TAM(1.00±0.10 vs 4.89±0.44)。
基金the National Natural Science Foundation of China,No.81072140,81172477the Medical Engineering Interdisciplinary Project of Shanghai Jiao Tong University,No.YG2013MS67,YG2014QN11~~
基金supported by a grant from the French National League against Cancer,Committee of the Marne.
文摘The JAR choriocarcinoma cell line share many of the characteristics of early placental trophoblast cells including the invasion properties. Matrix metallo-proteinases (MMPs), the main actors of matrix proteolysis, are involved in normal invasion as well as in the invasive character of tumor cells and the metastase formation. Tumor necrosis factor-α (TNF-α) is present in the placental environment and TNF-α levels are elevated in some placental pathologies. In the present work, we addressed whether TNF-α is a modulator of JAR cell MMP secretion. Following TNF-α stimulation, zymographic analysis showed the increased secretion of the active form of MMP-2 and to a lesser extent proMMP-2 and MMP-9. In addition, MMP-2 gene expression only increased slightly whereas MMP-9 and TIMP-1 transcripts were undetectable. This suggests that TNF-α may modulate the secretion of MMPs independently of MMP gene expression control.
