BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(X...BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(XIST)expression has been reported to be elevated in the serum of DN patients.AIM To evaluate the mechanism of lncRNA XIST in renal tubular epithelial cell(RTEC)pyroptosis in DN.METHODS A DN rat model was established through streptozotocin injection,and XIST was knocked down by tail vein injection of the lentivirus LV sh-XIST.Renal metabolic and biochemical indices were detected,and pathological changes in the renal tissue were assessed.The expression of indicators related to inflammation and pyroptosis was also detected.High glucose(HG)was used to treat HK2 cells,and cell viability and lactate dehydrogenase(LDH)activity were detected after silencing XIST.The subcellular localization and downstream mechanism of XIST were investigated.Finally,a rescue experiment was carried out to verify that XIST regulates NLR family pyrin domain containing 3(NLRP3)/caspase-1-mediated RTEC pyroptosis through the microRNA-15-5p(miR-15b-5p)/Toll-like receptor 4(TLR4)axis.RESULTS XIST was highly expressed in the DN models.XIST silencing improved renal metabolism and biochemical indices and mitigated renal injury.The expression of inflammation and pyroptosis indicators was significantly increased in DN rats and HG-treated HK2 cells;cell viability was decreased and LDH activity was increased after HGtreatment. Silencing XIST inhibited RTEC pyroptosis by inhibiting NLRP3/caspase-1. Mechanistically,XIST sponged miR-15b-5p to regulate TLR4. Silencing XIST inhibited TLR4 by promotingmiR-15b-5p. miR-15b-5p inhibition or TLR4 overexpression averted the inhibitory effect ofsilencing XIST on HG-induced RTEC pyroptosis.CONCLUSIONSilencing XIST inhibits TLR4 by upregulating miR-15b-5p and ultimately inhibits renal injury inDN by inhibiting NLRP3/caspase-1-mediated RTEC pyroptosis.展开更多
Background： Accumulating documents have demonstrated that long noncoding RNAs （lncRNAs） play critical roles in tumorigenesis. As an lncRNA, nuclear-enriched abundant transcript 1 （NEAT1） has been identified to be...Background： Accumulating documents have demonstrated that long noncoding RNAs （lncRNAs） play critical roles in tumorigenesis. As an lncRNA, nuclear-enriched abundant transcript 1 （NEAT1） has been identified to be involved in the progression of many types of cancers. However, the biological function of NE.4T1 in cervical cancer is not fully investigated. The aim of this study was to disclose the specific biological function of lncRNA NEATI in cervical cancer progression. Methods： Quantitative real-time polymerase chain reaction （qRT-PCR） was utilized to identify the expression of lncRNA NE,4 T1 in the cervical cancer tissues and cell lines. All cervical cancer samples used in this study were collected from the Affiliated Suzhou Hospital of Nanjing Medical University between September 2012 and September 2017. The correlation between NE,4T1 expression and the overall survival rate of cervical cancer patients was analyzed by Kaplan-Meier analysis. The effects of NEAT1 knockdown or overexpression on cell proliferation were tested by performing MTT assays and colony formation assays. Transwell assays were conducted to detect the migratory ability of cervical cancer cells, in which NEAT1 was silenced or overexpressed. Western blotting was utilized to validate whether NEAT1 promotes cervical cancer progression through activating PI3K-Akt signaling pathway. Results： High expression of NE,4T1 predicted poor prognosis of cervical cancer patients （χ^2= 0.735, P = 0.005）. Knockdown of NE,4T1 decreased the number of colonies in CaSki cell from 136.667 ± 13.503 to 71.667 ± 7.506 （t = -18.76, P = 0.003） and decreased the number of colonies in HeLa cell from 128.667 ± 13.317 to 65.667 ± 7.024 （t = -5.54, P = 0.031）. However, overexpression of NEA T1 increased the number of colonies in SiHa cell from 84.667 ± 12.014 to 150.667 ± 18.037 （t = 7.27, P = 0.018）. Knockdown of NEAT1 decreased the migratory number of CaSki cell from 100.333 ± 9.866 to 58.333 ± 5.859 （t = -8.08, P = 0.015） and reduced the migratory number in HeLa cell from 123.667± 12.097 to 67.667 ± 7.095 （t = -6.03, P = 0.026）. Overexpression of NEAT1 increased the migratory number of SiHa cell from 127.333 ±16.042 to 231.333 ±31.786 （t = 4.92, P = 0.039）. Conclusion： NEAT1 may exert oncogenic function in cervical cancer and serve as a novel therapeutic target for cervical cancer.展开更多
基金Supported by Natural Science Foundation of Shenzhen University General Hospital (SUGH2020QD011)
文摘BACKGROUND NLRP3-mediated pyroptosis is recognized as an essential modulator of renal disease pathology.Long noncoding RNAs(lncRNAs)are active participators of diabetic nephropathy(DN).X inactive specific transcript(XIST)expression has been reported to be elevated in the serum of DN patients.AIM To evaluate the mechanism of lncRNA XIST in renal tubular epithelial cell(RTEC)pyroptosis in DN.METHODS A DN rat model was established through streptozotocin injection,and XIST was knocked down by tail vein injection of the lentivirus LV sh-XIST.Renal metabolic and biochemical indices were detected,and pathological changes in the renal tissue were assessed.The expression of indicators related to inflammation and pyroptosis was also detected.High glucose(HG)was used to treat HK2 cells,and cell viability and lactate dehydrogenase(LDH)activity were detected after silencing XIST.The subcellular localization and downstream mechanism of XIST were investigated.Finally,a rescue experiment was carried out to verify that XIST regulates NLR family pyrin domain containing 3(NLRP3)/caspase-1-mediated RTEC pyroptosis through the microRNA-15-5p(miR-15b-5p)/Toll-like receptor 4(TLR4)axis.RESULTS XIST was highly expressed in the DN models.XIST silencing improved renal metabolism and biochemical indices and mitigated renal injury.The expression of inflammation and pyroptosis indicators was significantly increased in DN rats and HG-treated HK2 cells;cell viability was decreased and LDH activity was increased after HGtreatment. Silencing XIST inhibited RTEC pyroptosis by inhibiting NLRP3/caspase-1. Mechanistically,XIST sponged miR-15b-5p to regulate TLR4. Silencing XIST inhibited TLR4 by promotingmiR-15b-5p. miR-15b-5p inhibition or TLR4 overexpression averted the inhibitory effect ofsilencing XIST on HG-induced RTEC pyroptosis.CONCLUSIONSilencing XIST inhibits TLR4 by upregulating miR-15b-5p and ultimately inhibits renal injury inDN by inhibiting NLRP3/caspase-1-mediated RTEC pyroptosis.
基金This study was supported by grants from the National Natural Science Foundation of China （No. 81370719 and No. 81671535）, the Science Foundation of Jiangsu （No. BE2015642, and No. WSW023）, the Jiangsu Key Discipline of Human Assisted Reproduction Medicine Foundation （No. FXK201749）, and the Jiangsu Provincial Medical Youth Talent of the Project of Invigorating Health Care through Science, Technology, and Education （No. ZDRCA2016044）.
文摘Background： Accumulating documents have demonstrated that long noncoding RNAs （lncRNAs） play critical roles in tumorigenesis. As an lncRNA, nuclear-enriched abundant transcript 1 （NEAT1） has been identified to be involved in the progression of many types of cancers. However, the biological function of NE.4T1 in cervical cancer is not fully investigated. The aim of this study was to disclose the specific biological function of lncRNA NEATI in cervical cancer progression. Methods： Quantitative real-time polymerase chain reaction （qRT-PCR） was utilized to identify the expression of lncRNA NE,4 T1 in the cervical cancer tissues and cell lines. All cervical cancer samples used in this study were collected from the Affiliated Suzhou Hospital of Nanjing Medical University between September 2012 and September 2017. The correlation between NE,4T1 expression and the overall survival rate of cervical cancer patients was analyzed by Kaplan-Meier analysis. The effects of NEAT1 knockdown or overexpression on cell proliferation were tested by performing MTT assays and colony formation assays. Transwell assays were conducted to detect the migratory ability of cervical cancer cells, in which NEAT1 was silenced or overexpressed. Western blotting was utilized to validate whether NEAT1 promotes cervical cancer progression through activating PI3K-Akt signaling pathway. Results： High expression of NE,4T1 predicted poor prognosis of cervical cancer patients （χ^2= 0.735, P = 0.005）. Knockdown of NE,4T1 decreased the number of colonies in CaSki cell from 136.667 ± 13.503 to 71.667 ± 7.506 （t = -18.76, P = 0.003） and decreased the number of colonies in HeLa cell from 128.667 ± 13.317 to 65.667 ± 7.024 （t = -5.54, P = 0.031）. However, overexpression of NEA T1 increased the number of colonies in SiHa cell from 84.667 ± 12.014 to 150.667 ± 18.037 （t = 7.27, P = 0.018）. Knockdown of NEAT1 decreased the migratory number of CaSki cell from 100.333 ± 9.866 to 58.333 ± 5.859 （t = -8.08, P = 0.015） and reduced the migratory number in HeLa cell from 123.667± 12.097 to 67.667 ± 7.095 （t = -6.03, P = 0.026）. Overexpression of NEAT1 increased the migratory number of SiHa cell from 127.333 ±16.042 to 231.333 ±31.786 （t = 4.92, P = 0.039）. Conclusion： NEAT1 may exert oncogenic function in cervical cancer and serve as a novel therapeutic target for cervical cancer.
