Background： MicroRNAs （miRNAs） have been extensively studied over the decades and have been identified as potential molecular targets for cancer therapy. To date, many miRNAs have been found participating in the tu...Background： MicroRNAs （miRNAs） have been extensively studied over the decades and have been identified as potential molecular targets for cancer therapy. To date, many miRNAs have been found participating in the tumorigenesis of non-small cell lung cancer （NSCLC）. The present study was designed to evaluate the functions of miR-125b-1-3p in NSCLC cells. Methods： MiR-125b-1-3p expression was detected in tissue samples from 21 NSCLC patients and in NSCLC cell lines using the real-time polymerase chain reaction. A549 cell lines were transfected with a miR-125b-1-3p mimic or miR-125b-1-3p antisense. Cell counting kit-8, wound healing, Matrigel invasion assays, and flow cytometry were used to assess the effects of these transfections on cell growth, migration, invasion, and apoptosis, respectively. Western blotting was used to detect apoptosis-related proteins, expression ofS 1PR 1, and the phosphorylation status of STAT3. Significant differences between groups were estimated using Student＇s t-test or a one-way analysis of variance. Results： MiR-125b-1-3p was downregulated in NSCLC samples and cell lines. Overexpression ofmiR-125b-1-3p inhibited NSCLC cell proliferation （37.8 ± 9.1%, t = 3.191, P = 0.013）, migration （42.3 ± 6.7%, t = 6.321, P = 0.003）, and invasion （57.6 ±1 1.3%, t =4.112, P = 0.001） and simultaneously induced more NSCLC cell apoptosis （2.76 ± 0.78 folds, t = 3.772, P = 0.001）. MiR-125b-l-3p antisense resulted in completely opposite results. SIPRI was found as the target gene ofmiR-125b-1-3p. Overexpression of miR-125b-1-3p inhibited S I PRI protein expression （27.4 ± 6.1% of control, t = 4.083, P = 0.007）. In addition, SIPR1 siRNA decreased STAY3 phosphorylation （16.4 ±0.14% of control, t = 3.023, P = 0.015）, as in cells overexpressing miR-125b-1-3p （16.7 ± 0.17% of control, t = 4.162, P = 0.026）. Conclusion： Our results suggest that miR-125b-1-3p exerts antitumor functions in NSCLC cells by targeting SIPR1.展开更多
Curcumenol,an effective ingredient of Wenyujin,has been reported that exerted its antitumor potential in a few cancer types.However,the effect and molecular mechanism of curcumenol in lung cancer are largely unknown.H...Curcumenol,an effective ingredient of Wenyujin,has been reported that exerted its antitumor potential in a few cancer types.However,the effect and molecular mechanism of curcumenol in lung cancer are largely unknown.Here,we found that curcumenol induced cell death and suppressed cell proliferation in lung cancer cells.Next,we demonstrated that ferroptosis was the predominant method that contributed to curcumenol-induced cell death of lung cancer in vitro and vivo for the first time.Subsequently,using RNA sequencing,we found that the long non-coding RNA H19(lncRNA H19)was significantly downregulated in lung cancer cells treated with curcumenol,when compared to untreated controls.Overexpression of lncRNA H19 eliminated the anticancer effect of curcumenol,while lncRNA H19 knockdown promoted ferroptosis induced by curcumenol treatment.Mechanistically,we showed that lncRNA H19 functioned as a competing endogenous RNA to bind to miR-19b-3p,thereby enhanced the transcription activity of its endogenous target,ferritin heavy chain 1(FTH1),a marker of ferroptosis.In conclusion,our data show that the natural product curcumenol exerted its antitumor effects on lung cancer by triggering ferroptosis,and the lncRNA H19/miR-19b-3p/FTH1 axis plays an essential role in curcumenol-induced ferroptotic cell death.Therefore,our findings will hopefully provide a valuable drug for treating lung cancer patients.展开更多
文摘目的 探讨长链非编码RNA(long non-coding RNA,LncRNA)核富集转录本1(nuclear enriched abundant transcript 1,NEAT1)通过调节微小核糖核酸(micro RNAs,miR)-125b-5p/胰岛素样生长因子结合蛋白5(insulinlike growth factor binding protein 5,IGFBP5)轴对血管瘤内皮细胞增殖、凋亡和迁移的影响。方法 实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)、蛋白免疫印迹(Western blot)分别检测血管瘤组织(2016年3月~2019年3月收集,n=18)、瘤旁组织样本(2016年3月~2019年3月收集,n=18)以及人脐静脉内皮细胞HUVES,人血管瘤内皮细胞HemECs,HDEC中NEAT1,miR-125b-5p及IGFBP5蛋白表达。构建沉默NEAT1,同时沉默NEAT1和miR-125b-5p的HemECs细胞系,通过细胞活力检测试剂盒(cell counting kit-8,CCK-8)、台盼蓝染色、流式细胞术、划痕愈合实验、Western blot分别观察NEAT1和miR-125b-5p对HemECs细胞增殖、凋亡、迁移及IGFBP5,增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、B细胞淋巴瘤/白血病-2(B cell lymphoma/lewkmia-2,Bcl-2)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白表达的影响;双荧光素酶报告基因实验检测NEAT1与miR-125b-5p,mi R-125b-5p与IGFBP5的关系。结果 与瘤旁组织比较,血管瘤组织中NEAT1(2.87±0.22 vs 1.00±0.00),IGFBP5蛋白(1.45±0.14 vs 0.27±0.02)表达水平升高,miR-125b-5p(0.24±0.02 vs 1.00±0.00)表达水平降低,差异具有统计学意义(t=35.400~161.220,均P <0.05);与HUVES细胞比较,HemECs,HDEC细胞中NEAT1(2.76±0.24,1.78±0.13 vs 1.00±0.00),IGFBP5蛋白(1.31±0.15,0.78±0.06 vs 0.24±0.02)表达升高,miR-125b-5p表达(0.19±0.02,0.45±0.04 vs 1.00±0.00)降低,差异具有统计学意义(t=17.320~99.204,14.697~33.680,均P<0.05),且HemECs细胞中NEAT1和IGFBP5蛋白表达量最高,miR-125b-5p表达量最低,因此,选取HemECs细胞为研究对象;与si-NC组比较,si-NEAT1组NEAT1(0.32±0.02 vs 1.01±0.12)表达、A值(0.45±0.04 vs 1.13±0.11)、细胞生长率(32.28%±2.79%vs 99.41%±0.22%)、划痕愈合率(20.33%±1.23%vs 49.24%±2.43%)及IGFBP5(0.41±0.04 vs 1.31±0.20),PCNA(0.36±0.04 vs 1.27±0.14),Bcl-2(0.48±0.04 vs 1.39±0.16)和MMP-9(0.21±0.02 vs 1.09±0.10)蛋白表达降低,mi R-125b-5p(1.87±0.15 vs 1.02±0.10)表达、细胞凋亡率(45.58%±3.34%vs 12.36%±1.07%)升高,差异具有统计学意义(t=10.809~58.755,均P <0.05);下调miR-125b-5p减弱了沉默NEAT1对HemECs细胞增殖、迁移的抑制及对细胞凋亡的促进作用(t=9.218~15.010,均P <0.05);NEAT1与miR-125b-5p,miR-125b-5p与IGFBP5存在靶向调控关系。结论 沉默NEAT1通过上调miR-125b-5p来抑制IGFBP5表达,从而抑制HemECs细胞增殖、迁移,并促进细胞凋亡。
文摘Background： MicroRNAs （miRNAs） have been extensively studied over the decades and have been identified as potential molecular targets for cancer therapy. To date, many miRNAs have been found participating in the tumorigenesis of non-small cell lung cancer （NSCLC）. The present study was designed to evaluate the functions of miR-125b-1-3p in NSCLC cells. Methods： MiR-125b-1-3p expression was detected in tissue samples from 21 NSCLC patients and in NSCLC cell lines using the real-time polymerase chain reaction. A549 cell lines were transfected with a miR-125b-1-3p mimic or miR-125b-1-3p antisense. Cell counting kit-8, wound healing, Matrigel invasion assays, and flow cytometry were used to assess the effects of these transfections on cell growth, migration, invasion, and apoptosis, respectively. Western blotting was used to detect apoptosis-related proteins, expression ofS 1PR 1, and the phosphorylation status of STAT3. Significant differences between groups were estimated using Student＇s t-test or a one-way analysis of variance. Results： MiR-125b-1-3p was downregulated in NSCLC samples and cell lines. Overexpression ofmiR-125b-1-3p inhibited NSCLC cell proliferation （37.8 ± 9.1%, t = 3.191, P = 0.013）, migration （42.3 ± 6.7%, t = 6.321, P = 0.003）, and invasion （57.6 ±1 1.3%, t =4.112, P = 0.001） and simultaneously induced more NSCLC cell apoptosis （2.76 ± 0.78 folds, t = 3.772, P = 0.001）. MiR-125b-l-3p antisense resulted in completely opposite results. SIPRI was found as the target gene ofmiR-125b-1-3p. Overexpression of miR-125b-1-3p inhibited S I PRI protein expression （27.4 ± 6.1% of control, t = 4.083, P = 0.007）. In addition, SIPR1 siRNA decreased STAY3 phosphorylation （16.4 ±0.14% of control, t = 3.023, P = 0.015）, as in cells overexpressing miR-125b-1-3p （16.7 ± 0.17% of control, t = 4.162, P = 0.026）. Conclusion： Our results suggest that miR-125b-1-3p exerts antitumor functions in NSCLC cells by targeting SIPR1.
基金This work was financially funded by the grants National Natural Science Foundation of China(No.81874380 and 82022075,to Xinbing Sui,81730108 and 81973635,to Tian Xie,82104207,to Xueni Sun)Zhejiang Provincial Natural Science Foundation of China for Distinguished Young Scholars(No.LR18H160001,to Xinbing Sui)+5 种基金the Science and Technology Development Fund,Macao SAR(No.130/2017/A3,0099/2018/A3 and 0098/2021/A2,to Qibiao Wu)Science and Technology Planning Project of Guangdong Province(2020B1212030008,to Qibiao Wu)Zhejiang Provincial Natural Science Foundation of China(No.LQ20H160013,Ting DuanLQ21H160038,to Jiao Feng)Zhejiang Province Science and Technology Project of TCM(No.2019ZZ016,to Xinbing Sui2021ZQ058,to Ruonan Zhang,China).
文摘Curcumenol,an effective ingredient of Wenyujin,has been reported that exerted its antitumor potential in a few cancer types.However,the effect and molecular mechanism of curcumenol in lung cancer are largely unknown.Here,we found that curcumenol induced cell death and suppressed cell proliferation in lung cancer cells.Next,we demonstrated that ferroptosis was the predominant method that contributed to curcumenol-induced cell death of lung cancer in vitro and vivo for the first time.Subsequently,using RNA sequencing,we found that the long non-coding RNA H19(lncRNA H19)was significantly downregulated in lung cancer cells treated with curcumenol,when compared to untreated controls.Overexpression of lncRNA H19 eliminated the anticancer effect of curcumenol,while lncRNA H19 knockdown promoted ferroptosis induced by curcumenol treatment.Mechanistically,we showed that lncRNA H19 functioned as a competing endogenous RNA to bind to miR-19b-3p,thereby enhanced the transcription activity of its endogenous target,ferritin heavy chain 1(FTH1),a marker of ferroptosis.In conclusion,our data show that the natural product curcumenol exerted its antitumor effects on lung cancer by triggering ferroptosis,and the lncRNA H19/miR-19b-3p/FTH1 axis plays an essential role in curcumenol-induced ferroptotic cell death.Therefore,our findings will hopefully provide a valuable drug for treating lung cancer patients.
