应喜慧, 廖建, 鞠梅, 黄小艳, 李娟. 长非编码RNA-p53诱导转录本通过靶向miR-1297调控高糖处理HK-2细胞的迁移和上皮间质转化[J]. 临床肾脏病杂志, 2022, 22(6): 484-491. DOI: 10.3969/j.issn.1671-2390.2022.06.007
    引用本文: 应喜慧, 廖建, 鞠梅, 黄小艳, 李娟. 长非编码RNA-p53诱导转录本通过靶向miR-1297调控高糖处理HK-2细胞的迁移和上皮间质转化[J]. 临床肾脏病杂志, 2022, 22(6): 484-491. DOI: 10.3969/j.issn.1671-2390.2022.06.007
    Ying Xi-hui, Liao Jian, Ju Mei, Huang Xiao-yan, Li Juan. LINC-PINT regulated the migration and epithelial-mesenchymal transition of HK-2 cells treated with high glucose by targeting miR-1297[J]. Journal of Clinical Nephrology, 2022, 22(6): 484-491. DOI: 10.3969/j.issn.1671-2390.2022.06.007
    Citation: Ying Xi-hui, Liao Jian, Ju Mei, Huang Xiao-yan, Li Juan. LINC-PINT regulated the migration and epithelial-mesenchymal transition of HK-2 cells treated with high glucose by targeting miR-1297[J]. Journal of Clinical Nephrology, 2022, 22(6): 484-491. DOI: 10.3969/j.issn.1671-2390.2022.06.007

    长非编码RNA-p53诱导转录本通过靶向miR-1297调控高糖处理HK-2细胞的迁移和上皮间质转化

    LINC-PINT regulated the migration and epithelial-mesenchymal transition of HK-2 cells treated with high glucose by targeting miR-1297

    • 摘要: 目的 探讨长非编码RNA-p53诱导转录本(long intergenic non-protein coding RNAp53 induced transcript,LINC-PINT)对高糖(high glucose,HG)处理人肾小管上皮细胞HK-2迁移和上皮间质转化(epithelial-mesenchymal transition,EMT)的影响及其分子机制。方法 分别用5.5mmol/L、30 mmol/L葡萄糖处理HK-2细胞,记为正常糖(normal glucose,NG)组、高糖(high glu-cose,HG)组。将HG组HK-2细胞分为HG+si-NC组、HG+si-LINC-PINT组、HG+miR-NC组、HG+miR-1297组、HG+si-LINC-PINT+anti-miR-NC组、HG+si-LINC-PINT+anti-miR-1297组。RT-qPCR检测LINC-PINT和miR-1297的表达水平,采用Transwell和Western blot检测HK-2细胞迁移和相关蛋白的表达,双荧光素酶报告实验鉴定LINC-PINT和miR-1297的靶向关系。结果 与NG组比较,HG组HK-2细胞中,LINC-PINT(1.00±0.05)比(2.87±0.19)和miR-1297(1.02±0.07)比(0.43±0.04)的表达水平分别显著升高和降低,迁移细胞数(89.37±4.56)比(212.35±13.28)以及基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)(0.31±0.02)比(0.79±0.07)、Vimentin(0.22±0.01)比(0.58±0.03)、α-平滑肌肌动蛋白(α-smooth actin,α-SMA)(0.19±0.02)比(0.53±0.05)和纤维粘连蛋白(fibrimine,FN)(0.12±0.01)比(0.34±0.04)的表达水平显著升高,E-钙黏蛋白(E-cadherin)(0.69±0.07)比(0.27±0.02)的表达水平显著降低(P<0.05)。抑制LINC-PINT表达或过表达miR-1297显著降低迁移细胞数(197.28±10.25)比(137.53±7.59),(216.58±13.87)比(145.29±6.82)、MMP-2(0.83±0.05)比(0.47±0.04),(0.78±0.05)比(0.42±0.04)、波形蛋白(0.62±0.05)比(0.36±0.02),(0.56±0.04)比(0.32±0.03)、α-SMA(0.51±0.03)比(0.28±0.02),(0.56±0.03)比(0.30±0.02)和FN(0.37±0.03)比(0.23±0.03),(0.36±0.03)比(0.25±0.02)的表达水平,显著升高E-cadherin(0.24±0.02)比(0.58±0.03),(0.23±0.02)比(0.55±0.04)的表达水平(P<0.05)。LINC-PINT靶向调控miR-1297的表达(P<0.05)。下调miR-1297逆转了抑制LINC-PINT表达对HG处理HK-2细胞迁移和EMT的影响(P<0.05)。结论 LINC-PINT通过靶向miR-1297调控HG处理HK-2细胞的迁移和EMT。

       

      Abstract: Objective To explore the effect of long intergenic non-protein coding RNA-p53 induced transcript (LINC-PINT) on the migration and epithelial-mesenchymal transition (EMT) of human renal tubular epithelial HK-2 cells treated with high glucose (HG) and elucidate its molecular mechanism. Methods HK-2 cells were treated with 5.5 and 30 mmol/L glucose and recorded as NG and HG groups. HK-2 cells in HG group were divided into the groups of HG+si-NC,HG+si-LINC-PINT, HG+miR-NC,HG+miR-1297,HG+si-LINC-PINT+anti-miR-NC and HG+si-LINC-PINT+antimiR-1297. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used for detecting the expression levels of LINC-PINT and miR-1297. Transwell and Western blot for detecting the migration of HK-2 cells and the expression of related proteins. Dual luciferase reporter assay for examining the targeting relationship between LINC-PINT and miR-1297. Results Compared with NG group,the expression levels of LINC-PINT (1.00±0.05) vs (2.87±0.19) and miR-1297 (1.02±0.07) vs (0.43±0.04) significantly spiked and dropped obviously in HG group;number of migrated cells (89.37±4.56) vs (212.35±13.28) and expression levels of MMP-2 (0.31±0.02) vs (0.79±0.07),vimentin (0.22±0.01) vs (0.58±0.03),α-SMA (0.19±0.02) vs (0.53±0.05) and FN (0.12±0.01) vs (0.34±0.04) rose obviously while expression level of E-cadherin declined markedly (0.69±0.07) vs (0.27±0.02),P<0.05. Suppression of LINC-PINT expression or overexpression of miR-1297 significantly lowered the number of migrated cells (197.28±10.25) vs (137.53±7.59), (216.58±13.87) vs (145.29±6.82) and the expression levels of MMP-2 (0.83±0.05) vs (0.47±0.04), (0.78±0.05) vs (0.42±0.04),vimentin (0.62±0.05) vs (0.36±0.02), (0.56±0.04) vs (0.32±0.03),α-SMA (0.51±0.03) vs (0.28±0.02), (0.56±0.03) vs (0.30±0.02) and Fn (0.37±0.03) vs (0.23±0.03), (0.36±0.03) vs (0.25±0.02) and significantly boosted the expression level of E-cadherin (0.24±0.02) vs (0.58±0.03), (0.23±0.02) vs (0.55±0.04) (P<0.05). LINC-PINT targeted the expression of miR-1297 (P<0.05). A down-regulation of miR-1297 reversed the effect of suppressing the expression of LINC-PINT on the migration and EMT of HK-2 cells treated with HG (P<0.05). Conclusion LINC-PINT regulates the migration and EMT of HK-2 cells treated with HG through targeting miR-1297.

       

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