乳源免疫调节肽通过泛素-蛋白酶体途径抑制卵巢癌耐药性及其机制研究
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摘要
目的研究乳源免疫调节肽(PGPIPN)抑制卵巢癌耐药性及其机制。方法 MTT法检测PGPIPN与抗癌药顺铂(DDP)联合用药抑制人卵巢癌敏感细胞株(SKOV_3)、耐药细胞株(SKOV_3-DDP)和原代卵巢癌细胞的半数抑制浓度(IC_(50))和细胞增殖抑制率,PCR法和Western blot法检测人卵巢癌细胞株和原代卵巢癌细胞用药情况下泛素-蛋白酶体途径相关基因的表达。结果 PGPIPN和DDP联合用药细胞增殖抑制率显著高于单独DDP组或PGPIPN组,差异有统计学意义(P<0.05),PCR和Western blot结果表明PGPIPN通过调节泛素-蛋白酶体途径相关基因表达降低卵巢癌细胞的耐药性。PGPIPN促进SIAH和PSMA1表达,降低β-catenin基因的表达(P<0.05,P<0.01),且有剂量依赖性。结论 PGPIPN通过泛素-蛋白酶体途径降低卵巢癌细胞对DDP的耐药性。
Objective To study the inhibitory effect of immunomodulating peptide(PGPIPN) on the drug resistance and its mechanism in ovarian cancer.Methods The IC_(50) and cell proliferation inhibition rates were detected in human ovarian cancer cell lines(both sensitive cell line,SKOV3,and drug-resistant cell line,SKOV3-DDP)and primary ovarian cancer cell through the combination of the PGPIPN and cisplatin(DDP) by MTT method.The PCR and Western blot were used to detect the expressions of genes related to the ubiquitin-proteasome pathway in human ovarian cancer cell lines and primary ovarian cancer cells.Results The experimental results showed that the proliferation inhibition rate in combined drug group was higher than that in the groups treated only with DDP and PGPIPN and the difference was statistically significant(P<0.05).The results of PCR and Western blot indicated that PGPIPN lessened the resistance of ovarian cancer cells to DDP by regulating the expressions of genes related to the ubiquitin-proteasome pathway in human ovarian cancer cell lines and primary ovarian cancer cells.PGPIPN promoted the expression of SIAH and PSMA1 genes and reduced the expression of β-catenin gene(P<0.05,P<0.01) with a dose-dependent manner.Conclusion PGPIPN reduces the resistance of ovarian cancer cells to DDP through the ubiquitin proteasome pathway.
Objective To study the inhibitory effect of immunomodulating peptide(PGPIPN) on the drug resistance and its mechanism in ovarian cancer.Methods The IC_(50) and cell proliferation inhibition rates were detected in human ovarian cancer cell lines(both sensitive cell line,SKOV3,and drug-resistant cell line,SKOV3-DDP)and primary ovarian cancer cell through the combination of the PGPIPN and cisplatin(DDP) by MTT method.The PCR and Western blot were used to detect the expressions of genes related to the ubiquitin-proteasome pathway in human ovarian cancer cell lines and primary ovarian cancer cells.Results The experimental results showed that the proliferation inhibition rate in combined drug group was higher than that in the groups treated only with DDP and PGPIPN and the difference was statistically significant(P<0.05).The results of PCR and Western blot indicated that PGPIPN lessened the resistance of ovarian cancer cells to DDP by regulating the expressions of genes related to the ubiquitin-proteasome pathway in human ovarian cancer cell lines and primary ovarian cancer cells.PGPIPN promoted the expression of SIAH and PSMA1 genes and reduced the expression of β-catenin gene(P<0.05,P<0.01) with a dose-dependent manner.Conclusion PGPIPN reduces the resistance of ovarian cancer cells to DDP through the ubiquitin proteasome pathway.
引文
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[2]周艳,秦宜德,董华胜,等.Β-CM-7对MCF-7、SKOV3和SK-N-SH肿瘤细胞增殖和凋亡的影响[J].安徽医科大学学报,2008,43(1):5-9.
[3]顾芳,秦宜德,董华胜,等.乳源免疫调节肽对小鼠抗氧化和抗疲劳作用研究[J].营养学报,2006,28(4):326-8.
[4]李素萍,秦宜德,董琼珠,等.饲喂乳源免疫调节肽对大鼠生长和免疫的影响[J].安徽医科大学学报,2005,40(6):499-501.
[5]魏彩,秦宜德,郑欣,等.乳源免疫调节肽体外抑制人卵巢癌细胞侵袭和转移[J].中国药理学通报,2013,29(1):42-8.
[6]郑欣,何晓光,张文晓,等.免疫调节肽对卵巢癌SKOV3细胞凋亡及相关基因表达的影响[J].安徽医科大学学报,2012,47(7):758-62.
[7]周娟,刘琛,顾芳,赵梦静,等.乳源免疫调节肽提高DDP对卵巢癌细胞的影响[J].安徽医科大学学报,2016,51(10):1440-44.
[8]Toerres-Llanez,Mde J,Vallejo-Cordoba B,González-Córdova A F.Bioactive peptides derived from milk proteins[J].Arch Latinoam Nutr,2005,55(2):111-7.
[9]Baldi A,Ioannis P,Chiara P,et al.Biological effects of milk proteins and their peptide with emphasis on those related to the gastrointestinal ecosystem[J].J Dairy Res,2005,72:66-72.
[10]李小勤,筛选和鉴定抗卵巢癌活性六肽的靶点蛋白[D].合肥:安徽医科大学,2014.
[11]Wu H,Shi Y,Lin Y,et al.Eukaryotic translation elongation factor 1 delta inhibits the ubiquitin ligase activity of SIAH-1[J].Mol Cell Biochem,2011,357(1-2):209-15.
[12]Matsuzawa S,Li C,Ni C Z,et al.Structural analysis of Siah1and its interactions with Siah-interacting protein(SIP)[J].J Biol Chem,2003,278(3):1837-40.