[目的]探究miR-325-3p靶向PRDX4对肾细胞癌细胞增殖、侵袭及凋亡的影响。[方法]设肾细胞癌细胞Caki-1组、miR-NC组、miR-325-3p-mimics组(过表达)、miR-325-3p-inhibitor组(低表达),测定各组细胞增殖、单克隆形成数目、凋亡率、侵袭水...[目的]探究miR-325-3p靶向PRDX4对肾细胞癌细胞增殖、侵袭及凋亡的影响。[方法]设肾细胞癌细胞Caki-1组、miR-NC组、miR-325-3p-mimics组(过表达)、miR-325-3p-inhibitor组(低表达),测定各组细胞增殖、单克隆形成数目、凋亡率、侵袭水平以及miR-325-3p、PRDX4水平。[结果]miR-325-3p-inhibitor组OD值(0.93±0.03)、存活率(86.58±6.36)%、单克隆形成数目(1062.29±102.78)、穿膜数(1917.34±425.35)、PRDX4 mRNA和蛋白表达水平(4.63±0.28、1.82±0.18)高于miR-325-3p-mimics组[(0.42±0.02)、(42.25±7.20)%、(239.89±35.27)个、(293.85±95.28)个、(2.04±0.24)、(0.38±0.07)](P<0.05),细胞凋亡率(1.12±0.29)%、miR-325-3p表达水平(1.42±0.38)低于miR-325-3p-mimics组(7.14±1.11)%、(5.68±0.37)(P<0.05)。[结论]miR-325-3p上调可以抑制肾细胞癌细胞的增殖(76.59%±7.30%vs 42.25%±7.20%)、迁移侵袭(702.28±111.52 vs 293.85±95.28),同时诱导细胞凋亡(3.46±1.04 vs 7.14±1.11),而这些过程主要是通过miR-325-3p与PRDX4的相互作用实现的。展开更多
Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chro...Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chronic pain conditions. However, the role of mi RNA-325-5 p in chronic visceral pain remains unknown. The present study was designed to determine the roles and mechanism of mi RNA-325-5 p in a rat model of chronic visceral pain.This model was induced by neonatal colonic inflammation(NCI). In adulthood, NCI led to a significant reduction in the expression of mi RNA-325-5 p in colon-related dorsal root ganglia(DRGs), starting to decrease at the age of4 weeks and being maintained to 8 weeks. Intrathecal administration of mi RNA-325-5 p agomir significantly enhanced the colorectal distention(CRD) threshold in a time-dependent manner. NCI also markedly increased the expression of CCL2(C-C motif chemokine ligand 2) in colon-related DRGs at the m RNA and protein levels relative to age-matched control rats. The expression of CXCL12, IL33, SFRS7, and LGI1 was not significantlyaltered in NCI rats. CCL2 was co-expressed in Neu Npositive DRG neurons but not in glutamine synthetasepositive glial cells. Furthermore, CCL2 was mainly expressed in isolectin B4-binding-and calcitonin generelated peptide-positive DRG neurons but in few NF-200-positive cells. More importantly, CCL2 was expressed in mi R-325-5 p-positive DRG neurons. Intrathecal injection of mi RNA-325-5 p agomir remarkably reduced the upregulation of CCL2 in NCI rats. Administration of Bindarit, an inhibitor of CCL2, markedly raised the CRD threshold in NCI rats in a dose-and time-dependent manner. These data suggest that NCI suppresses mi RNA-325-5 p expression and enhances CCL2 expression, thus contributing to visceral hypersensitivity in adult rats.展开更多
文摘[目的]探究miR-325-3p靶向PRDX4对肾细胞癌细胞增殖、侵袭及凋亡的影响。[方法]设肾细胞癌细胞Caki-1组、miR-NC组、miR-325-3p-mimics组(过表达)、miR-325-3p-inhibitor组(低表达),测定各组细胞增殖、单克隆形成数目、凋亡率、侵袭水平以及miR-325-3p、PRDX4水平。[结果]miR-325-3p-inhibitor组OD值(0.93±0.03)、存活率(86.58±6.36)%、单克隆形成数目(1062.29±102.78)、穿膜数(1917.34±425.35)、PRDX4 mRNA和蛋白表达水平(4.63±0.28、1.82±0.18)高于miR-325-3p-mimics组[(0.42±0.02)、(42.25±7.20)%、(239.89±35.27)个、(293.85±95.28)个、(2.04±0.24)、(0.38±0.07)](P<0.05),细胞凋亡率(1.12±0.29)%、miR-325-3p表达水平(1.42±0.38)低于miR-325-3p-mimics组(7.14±1.11)%、(5.68±0.37)(P<0.05)。[结论]miR-325-3p上调可以抑制肾细胞癌细胞的增殖(76.59%±7.30%vs 42.25%±7.20%)、迁移侵袭(702.28±111.52 vs 293.85±95.28),同时诱导细胞凋亡(3.46±1.04 vs 7.14±1.11),而这些过程主要是通过miR-325-3p与PRDX4的相互作用实现的。
基金supported by grants from the National Natural Science Foundation of China (81471137, 31730040, and 81771187)the Priority Academic Program Development of Jiangsu Higher Education Institutions of China
文摘Chronic visceral hypersensitivity is an important type of chronic pain with unknown etiology and pathophysiology. Recent studies have shown that epigenetic regulation plays an important role in the development of chronic pain conditions. However, the role of mi RNA-325-5 p in chronic visceral pain remains unknown. The present study was designed to determine the roles and mechanism of mi RNA-325-5 p in a rat model of chronic visceral pain.This model was induced by neonatal colonic inflammation(NCI). In adulthood, NCI led to a significant reduction in the expression of mi RNA-325-5 p in colon-related dorsal root ganglia(DRGs), starting to decrease at the age of4 weeks and being maintained to 8 weeks. Intrathecal administration of mi RNA-325-5 p agomir significantly enhanced the colorectal distention(CRD) threshold in a time-dependent manner. NCI also markedly increased the expression of CCL2(C-C motif chemokine ligand 2) in colon-related DRGs at the m RNA and protein levels relative to age-matched control rats. The expression of CXCL12, IL33, SFRS7, and LGI1 was not significantlyaltered in NCI rats. CCL2 was co-expressed in Neu Npositive DRG neurons but not in glutamine synthetasepositive glial cells. Furthermore, CCL2 was mainly expressed in isolectin B4-binding-and calcitonin generelated peptide-positive DRG neurons but in few NF-200-positive cells. More importantly, CCL2 was expressed in mi R-325-5 p-positive DRG neurons. Intrathecal injection of mi RNA-325-5 p agomir remarkably reduced the upregulation of CCL2 in NCI rats. Administration of Bindarit, an inhibitor of CCL2, markedly raised the CRD threshold in NCI rats in a dose-and time-dependent manner. These data suggest that NCI suppresses mi RNA-325-5 p expression and enhances CCL2 expression, thus contributing to visceral hypersensitivity in adult rats.
