目的 探讨维立西呱对脂多糖(LPS)诱导的大鼠心肌细胞炎症反应和细胞凋亡的影响以及其对环状RNA PRKCI(circ PRKCI)表达的调控机制。方法 取大鼠心肌细胞H9C2,采用LPS诱导方式构建心肌细胞损伤模型;按随机数字表法分为对照组、LPS组、LPS...目的 探讨维立西呱对脂多糖(LPS)诱导的大鼠心肌细胞炎症反应和细胞凋亡的影响以及其对环状RNA PRKCI(circ PRKCI)表达的调控机制。方法 取大鼠心肌细胞H9C2,采用LPS诱导方式构建心肌细胞损伤模型;按随机数字表法分为对照组、LPS组、LPS+维立西呱-L组(1μmol/L)、LPS+维立西呱-M组(3μmol/L)、LPS+维立西呱-H组(10μmol/L)、LPS+空载体质粒(p c DNA)组、LPS+c irc PRKCI过表达载体(p c DNA-c irc PRKCI)组、LPS+维立西呱-H+阴性对照(s h-NC)组、LPS+维立西呱-H+circ PRKCI慢病毒短发夹RNA(sh-circ PRKCI)组。检测并比较各组H9C2细胞肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)等炎症因子水平,细胞凋亡率和裂解的胱天蛋白酶(Cleaved Caspase)-3、Cleaved Ca s p a s e-9等凋亡相关蛋白表达水平,以及c irc PRKCI mRNA表达水平。结果 与对照组比较,LPS组TNF-α水平、IL-6水平、细胞凋亡率及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显升高(均P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组上述指标均明显降低(均P<0.05),且LPS+维立西呱-L组>LPS+维立西呱-M组>LPS+维立西呱-H组(均P<0.05)。与对照组比较,LPS组c irc PRKCI mRNA表达水平明显降低(P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组c irc PRKCI mRNA表达水平均明显升高(均P<0.05),且LPS+维立西呱-L组<LPS+维立西呱-M组<LPS+维立西呱-H组(均P<0.05)。与LPS+pc DNA组比较,LPS+pc DNA-circ PRKCI组circ PRKCI mRNA表达水平明显升高(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显降低(均P<0.05)。与LPS+维立西呱-H+sh-NC组比较,LPS+维立西呱-H+sh-circ PRKCI组circ PRKCI mRNA表达水平明显降低(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平均明显升高(均P<0.05)。结论 维立西呱可抑制LPS诱导的大鼠心肌细胞炎症反应和细胞凋亡,其作用机制是上调circ PRKCI mRNA表达。展开更多
BACKGROUND The circular RNA circ-PRKCI is an endogenous non-coding RNA that forms a covalently closed ring after reverse splicing, which plays a key role in the occurrence and development of multiple digestive system ...BACKGROUND The circular RNA circ-PRKCI is an endogenous non-coding RNA that forms a covalently closed ring after reverse splicing, which plays a key role in the occurrence and development of multiple digestive system tumors.AIM To investigate the role and mechanism of circ-PRKCI in the occurrence and development of hepatocellular carcinoma(HCC).METHODS This study used real-time polymerase chain reaction to detect the expression of circ-PRKCI in tumor tissues, tumor adjacent tissues, and blood in patients with HCC and other digestive system tumor cells. A series of functional tests were performed to explore whether circ-PRKCI affects the growth of HCC cells and what is its mechanism in HCC. Meanwhile, fluorescence in situ hybridization was used to detect the subcellular localization of circ-PRKCI. Survival analysis was performed to predict the correlation between circ-PRKCI and the prognosis of HCC. Chi-square test and t-test were performed for statistical analyses.RESULTS The level of circ-PRKCI was significantly higher in HCC tissues than in tumor adjacent tissues, and in HCC cell lines than in cells lines of esophageal, liver,stomach, and colon cancers. A series of functional tests showed that circ-PRKCI substantially inhibited cell apoptosis and promoted cell invasion. It was foundthat circ-PRKCI can act as the sponge of miRNA-545 to reduce the expression of AKT3 protein. Moreover, the result of survival analysis showed that circ-PRKCI target gene E2 F7 can reduce liver cancer patients' survival rate. And clinical data suggested that the distribution of circ-PRKCI rose with the depth of invasion,lymph node metastasis, distant metastasis, and TNM stage, indicating that circPRKCI may affect the survival and prognosis of patients with HCC by regulating E2 E7.CONCLUSION This study explores the role and mechanism of circ-PRKCI in HCC, which provides a new research direction and theoretical basis for the treatment of HCC.展开更多
文摘目的 探讨维立西呱对脂多糖(LPS)诱导的大鼠心肌细胞炎症反应和细胞凋亡的影响以及其对环状RNA PRKCI(circ PRKCI)表达的调控机制。方法 取大鼠心肌细胞H9C2,采用LPS诱导方式构建心肌细胞损伤模型;按随机数字表法分为对照组、LPS组、LPS+维立西呱-L组(1μmol/L)、LPS+维立西呱-M组(3μmol/L)、LPS+维立西呱-H组(10μmol/L)、LPS+空载体质粒(p c DNA)组、LPS+c irc PRKCI过表达载体(p c DNA-c irc PRKCI)组、LPS+维立西呱-H+阴性对照(s h-NC)组、LPS+维立西呱-H+circ PRKCI慢病毒短发夹RNA(sh-circ PRKCI)组。检测并比较各组H9C2细胞肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)等炎症因子水平,细胞凋亡率和裂解的胱天蛋白酶(Cleaved Caspase)-3、Cleaved Ca s p a s e-9等凋亡相关蛋白表达水平,以及c irc PRKCI mRNA表达水平。结果 与对照组比较,LPS组TNF-α水平、IL-6水平、细胞凋亡率及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显升高(均P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组上述指标均明显降低(均P<0.05),且LPS+维立西呱-L组>LPS+维立西呱-M组>LPS+维立西呱-H组(均P<0.05)。与对照组比较,LPS组c irc PRKCI mRNA表达水平明显降低(P<0.05);与LPS组比较,LPS+维立西呱-L组、LPS+维立西呱-M组、LPS+维立西呱-H组c irc PRKCI mRNA表达水平均明显升高(均P<0.05),且LPS+维立西呱-L组<LPS+维立西呱-M组<LPS+维立西呱-H组(均P<0.05)。与LPS+pc DNA组比较,LPS+pc DNA-circ PRKCI组circ PRKCI mRNA表达水平明显升高(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cle a ve d Ca s p a s e-3、Cle a ve d Ca s p a s e-9蛋白表达水平均明显降低(均P<0.05)。与LPS+维立西呱-H+sh-NC组比较,LPS+维立西呱-H+sh-circ PRKCI组circ PRKCI mRNA表达水平明显降低(P<0.05),而TNF-α水平、IL-6水平、细胞凋亡率以及凋亡相关蛋白Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平均明显升高(均P<0.05)。结论 维立西呱可抑制LPS诱导的大鼠心肌细胞炎症反应和细胞凋亡,其作用机制是上调circ PRKCI mRNA表达。
文摘BACKGROUND The circular RNA circ-PRKCI is an endogenous non-coding RNA that forms a covalently closed ring after reverse splicing, which plays a key role in the occurrence and development of multiple digestive system tumors.AIM To investigate the role and mechanism of circ-PRKCI in the occurrence and development of hepatocellular carcinoma(HCC).METHODS This study used real-time polymerase chain reaction to detect the expression of circ-PRKCI in tumor tissues, tumor adjacent tissues, and blood in patients with HCC and other digestive system tumor cells. A series of functional tests were performed to explore whether circ-PRKCI affects the growth of HCC cells and what is its mechanism in HCC. Meanwhile, fluorescence in situ hybridization was used to detect the subcellular localization of circ-PRKCI. Survival analysis was performed to predict the correlation between circ-PRKCI and the prognosis of HCC. Chi-square test and t-test were performed for statistical analyses.RESULTS The level of circ-PRKCI was significantly higher in HCC tissues than in tumor adjacent tissues, and in HCC cell lines than in cells lines of esophageal, liver,stomach, and colon cancers. A series of functional tests showed that circ-PRKCI substantially inhibited cell apoptosis and promoted cell invasion. It was foundthat circ-PRKCI can act as the sponge of miRNA-545 to reduce the expression of AKT3 protein. Moreover, the result of survival analysis showed that circ-PRKCI target gene E2 F7 can reduce liver cancer patients' survival rate. And clinical data suggested that the distribution of circ-PRKCI rose with the depth of invasion,lymph node metastasis, distant metastasis, and TNM stage, indicating that circPRKCI may affect the survival and prognosis of patients with HCC by regulating E2 E7.CONCLUSION This study explores the role and mechanism of circ-PRKCI in HCC, which provides a new research direction and theoretical basis for the treatment of HCC.
