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【Objective】 To explore the role of RYBP in activating PARP-1 dependent Parthanatos and promoting response to YM155 in esophageal squamous cell carcinoma. 【Methods】 CCK-8 and flow cytometry were used to analyze the inhibition ratio and cell death percentage after YM155 treatment in both RYBP overexpression group and control group. Western blotting was used to detect the expression of Parthanatos-related proteins. 【Results】 Compared with control group, RYBP overexpression group showed higher inhibition ratio and cell death percentage after YM155 treatment. Overexpression of RYBP activated PARP-1 with or without YM155 treatment. Besides, after YM155 treatment, KYSE170-RYBP showed more PAR accumulation in the nucleus, AIF translocation from mitochondria to the nucleus than control cells. 【Conclusion】 RYBP can activate PARP-1/PAR/AIF-dependent induced Parthanatos in esophageal squamous cell carcinoma and enhance response to YM155.
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Purpose To investigate the expression and clinical significance of RINGI and YYI binding protein (RYBP)in pancreatic carcinoma (PC).Methods RYBP expression was detected in 51 cases of PC tissues and paired adjacent nontumor tissues by immunohistochemistry.The correlation between RYBP expression and the survival time of PC patients after surgery was analyzed by Kaplan-Meier method.Results Compared to the expression in adjacent non-tumor tissues,RYBP was significantly lower expressed in PC tissues (P ＜ 0.05).In addition,RYBP expression had a significant correlation with tumor diameter and tumor staging of patients (P ＜ 0.05),but no significant correlation with other characteristics of patients,such as gender,age,smoking,alcohol intake,tumor number,metastasis or CA199 level (P ＞ 0.05).The survival analysis showed that the survival time of PC patients with positive expression of RYBP was significantly longer than patients with negative expression of RYBP (P ＜ 0.05).Conclusion RYBP is significantly lower expressed in PC tissues,which might be involved in the pathogenesis of PC.
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Objective To investigate the target genes in RYBP-mediated leukemia cell apoptosis by high-flux sequencing. Methods The HL-60 cell line with knockdown of RYBP was set up. MRNA and microRNA sequencing was conducted by the next-generation sequencing technology. Result MRNAs and miRNAs were dys-regulated in HL-60 cell line with knockdown of RYBP. The results of KEGG analysis indicated that the down-regu-lated genes were associated with cancer transcription regulation and amino acid metabolism. SPI1 and miR-214-3p were shown downregulated after knockdown of RYBP. Conclusion The target genes in RYBP-mediated leukemia cell apoptosis include BBC3,BAI1,SESN2 ,CCNG2,JAK3,STAT4,SPI1,BCL6,CD11b and hsa-miR-214-3p.
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Objective To establish stable HL-60 cell line with stable RYBP gene silencing using lentivirus-mediated RNA interference. Methods Five special shRNAs for RYBP gene were cloned to lentivirus vector.Recombinate lentivirus vectors were packed into lentivirus,which were used to infect HL-60 cells, and took empty vector and non-specific shRNA as control groups. Stable infected cells were selected with puromycin in 8 μg/ml concentration.The expression levels of RYBP were analyzed by Western blot,and confirmed the most effective RYBP shRNA.Then the level of mRNA was analyzed by real-time PCR.Results Stable infected cells were selected by puromycin successfully.Comparing to control groups,the expression of RYBP were reduced at different degrees (P ＜ 0.01). And RYBP shRNA2 took the most silencing effect, the RYBP mRNA was decreased by more than 95 ％ (P ＜ 0.05).Conclusion The shRNA2 targeting RYBP gene can effectively inhibit the expression of RYBP. HL-60 cell line with stable RYBP gene silencing were constructed successfully,which had provided experiment fundament for further studying the function of RYBP.