IP10-CDR3 Reduces The Viability And Induces The Apoptosis Of Ovarian Cancer Cells By Down-Regulating The Expression Of Bcl-2 And Caspase 3
IP10-CDR3 Reduces The Viability And Induces The Apoptosis Of Ovarian Cancer Cells By Down-Regulating The Expression Of Bcl-2 And Caspase 3
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IP10-CDR3通过下调Bcl-2和Caspase 3的表达降低卵巢癌细胞的活力并诱导其凋亡
DOI：
10.2147/ott.s209757
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发表时间：
2019-01-01
影响因子：
4
通讯作者：
He, Linsheng
中科院分区：
文献类型：
--
作者：
Chen, Qi;He, Quan;He, Linsheng
目的：探讨干扰素γ诱导蛋白10（IP10）和T细胞受体互补决定区3（CDR3）对卵巢癌细胞的影响及其机制。方法：将IP10和CDR3与单链抗体（scfv）和铜绿假单胞菌外毒素基因突变体（PE40）连接，构建IP10-CDR3scfv并IP10-CDR3-PE40scfv。然后，我们构建了pcDNA3.1-IP10-CDR3scfv和pcDNA3.1-IP10-CDR3-PE40scfv质粒，并通过HindIII/EcoRI酶切验证。将 SKOV3 细胞和 HOSEpiC 细胞与异硫氰酸荧光素 (FITC) 标记的 IP10-CDR3scfv 和 IP10-CDR3-PE40scfv 蛋白一起孵育，并通过流式细胞术检查蛋白水平。基因转染后，SKOV3细胞分为四组：Control、pcDNA3.1(+)阴性对照(NC)（pcDNA3.1(+) NC转染）、IP10-CDR3scfv（IP10-CDR3scfv转染）和IP10-CDR3-PE40scfv（IP10-CDR3-PE40scfv转染）。通过RT-PCR和Western blot测定IP10、CDR3、Caspase-3、裂解的Caspase-3和Bcl-2的水平。分别采用CCK-8法和Annexin V-FITC/PI法检测细胞活力和凋亡情况。结果：SKOV3+IP10-CDR3scfv组和SKOV3+IP10-CDR3-PE40scfv组FITC标记的IP10-CDR3scfv和IP10-CDR3-PE40scfv蛋白水平显着高于SKOV3组。组（P
Purpose: This study aimed to explore the effects of interferon-gamma inducible protein 10 (IP10) and complementarity-determining region 3 (CDR3) of T cells receptor on ovarian cancer cells and the involved mechanisms.Methods: IP10 and CDR3 were linked with single-chain antibody (scfv) and exotoxin gene muton of Pseudomonas aeruginosa (PE40) to construct IP10-CDR3scfv and IP10-CDR3-PE40scfv. Then, we constructed pcDNA3.1-IP10-CDR3scfv and pcDNA3.1-IP10-CDR3-PE40scfv plasmids which were proved by HindIII/EcoRI digestion. SKOV3 cells and HOSEpiC cells were incubated with fluorescein isothiocyanate (FITC) labeled IP10-CDR3scfv and IP10-CDR3-PE40scfv proteins and protein levels were examined by flow cytometry. After gene transfection, SKOV3 cells were divided into four groups: Control, pcDNA3.1(+) negative control (NC) (pcDNA3.1(+) NC transfection), IP10-CDR3scfv (IP10-CDR3scfv transfection) and IP10-CDR3-PE40scfv (IP10-CDR3-PE40scfv transfection). Levels of IP10, CDR3, Caspase-3, cleaved Caspase-3 and Bcl-2 were determined by RT-PCR and Western blot. Cell viability and apoptosis were investigated by CCK-8 assay and Annexin V-FITC/PI assay, respectively.Results: The levels of FITC-labeled IP10-CDR3scfv and IP10-CDR3-PE40scfv proteins in the SKOV3+IP10-CDR3scfv group and the SKOV3+IP10-CDR3-PE40scfv group were remarkably higher than that in the SKOV3 group (P