ZFX promotes tumorigenesis and confers chemotherapy resistance in esophageal squamous cell carcinoma

IntroductionZinc finger X-chromosomal protein (ZFX) has been shown to be essential for the development and progression of multiple types of human cancers. However, its potential roles in esophageal squamous cell carcinoma (ESCC) have not yet been elucidated.Materials and methodsEighty-three pairs of frozen ESCC samples and their para-cancer samples and 24 fresh ESCC samples were collected. In vitro chemosensitivity was tested using the histoculture drug response assay. Quantitative RT-PCR and western blotting were used to measure the expression of functional genes. The effects of ZFX on cell growth, cell apoptosis, and chemosensitivity of the esophageal cancer cells were assessed.ResultsWe found that ZFX was more upregulated in ESCC tissues than in the para-cancer tissues, and its high expression was correlated with inferior clinicopathological characteristics and overall survival. Multivariate analysis revealed that ZFX was an independent prognostic factor in ESCC patients. In ESCC cell lines, ZFX silencing suppressed cell growth and induced cell apoptosis. In addition, ZFX expression was negatively correlated with the sensitivity of fresh ESCC tissues to chemotherapeutic drugs, including cisplatin, docetaxel, fluorouracil, and irinotecan. Furthermore, the depletion of ZFX sensitized ESCC cells to cisplatin, and docetaxel treatment. Mechanistically, ZFX silencing resulted in the inactivation of the MEK/ERK pathway, which mediated the downregulation of P-glycoprotein expression.ConclusionOur study therefore indicates that ZFX possibly plays a critical role in ESCC tumorigenesis and chemotherapy resistance and could be a significant prognostic biomarker and therapeutic target for ESCC.     

Zinc finger protein x-linked (ZFX) contributes to patient prognosis,cell proliferation and apoptosis in human laryngeal squamous cell carcinoma

Zinc finger protein, X-linked (ZFX) gene locus on the human X chromosome is structurally similar to the zinc finger protein, Y-linked gene. Its role in human laryngeal squamous cell carcinoma (LSCC) is still not clearly defined. This study was focused on investigating the role of zinc-finger protein X-linked (ZFX) in human LSCC. Expression levels of ZFX were examined in LSCC tissues, corresponding adjacent non-tumoral tissues and vocal leukoplakia tissues by immunohistochemistry (IHC). The association with the expression level of ZFX and LSCC clincopathological parameters was analyzed. The prognostic value of ZFX expression was also analyzed. Lentivirus-mediated RNA interference was applied to silence ZFX expression and the effects of ZFX knockdown on the growth of human LSCC primary cells was investigated. Overexpression of ZFX was found in LSCC tissues. The expression of ZFX was associated with the clinical stage of LSCC. Patients with higher level of ZFX experienced a poorer prognosis compared to those with lower level of ZFX. Knockdown of ZFX inhibited cell proliferation, colony formation and migration of LSCC primary cells. Moreover, ZFX silencing induced cell apoptosis. These results provide the convincing evidence for the first time that ZFX plays an important role in LSCC development and could be a potential therapeutic target or prognostic predictor for LSCC.     

黄芩素对人胆囊癌细胞系SGC996细胞活力及ZFX蛋白表达的干预作用

目的：探讨中药黄芩提取物黄芩素对人胆囊癌的生物效应及机制。方法：将人胆囊癌细胞系SGC996用黄芩素处理48小时,采用MTT法检测治疗后SGC996的细胞活力；通过对透过transwell膜的SGC996细胞进行计数评估黄芩素对胆囊癌侵袭转移能力的影响；通过流式细胞术检测黄芩素处理后,SGC996细胞的凋亡情况并用western blot方法检测细胞锌指X染色体蛋白(zinc finger X-chromosomal protein,ZFX)的表达；结果：对照组,10μmol/L黄芩素组,20μmol/L黄芩素组,40μmol/L黄芩素组,80μmol/L黄芩素组,160μmol/L黄芩素组,320μmol/L黄芩素组对SGC996细胞活力的抑制率分别为0,0.4±0.9,4.2±1.5,19.3±3.8,37.9±5.8,61.6±7.8,84.2±10.2 (p<0.05)。对照组,40μmol/L黄芩素组,80μmol/L黄芩素组,160μmol/L黄芩素组对SGC996细胞的凋亡诱导率分别为0.5±0.5,7.5±1.2,16.3±1.9,31.2±2.8 (p<0.05)；对SGC996细胞转移抑制率分别为0,25.6±6.6,57.3±7.9,84.1±11.9 (p<0.05)。Western blot结果显示黄芩素对ZFX有显著的抑制作用。结论： 黄芩素有良好的抗神经胆囊癌的生物活性,其抗肿瘤效应的机制可能和下调ZFX蛋白有关。     

黄芩素对人胆囊癌细胞系SGC996细胞活力及细胞锌指X染色体蛋白表达的干预作用

目的探讨中药黄芩提取物黄芩素对人胆囊癌的生物效应及机制。方法将人胆囊癌细胞系SGC996用黄芩素处理48h,采用MTT法检测治疗后SGC996的细胞活力;通过对透过transwell膜的SGC996细胞进行计数,评估黄芩素对胆囊癌侵袭转移能力的影响;通过流式细胞术检测黄芩素处理后,SGC996细胞的凋亡情况,并用Western blot方法检测细胞锌指X染色体蛋白(ZFX)的表达。结果 40、80、160、320μmol/L黄芩素组对SGC996细胞活力的抑制率与对照组比较,差异有统计学意义[(19.3±3.8)%、(37.9±5.8)%、(61.6±7.8)%、(84.2±10.2)%比0,P0.05]。40、80、160μmol/L黄芩素组对SGC996细胞的凋亡诱导率与对照组比较,差异有统计学意义[(7.5±1.2)%、(16.3±1.9)%、(31.2±2.8)%比(0.5±0.5)%,P0.05]。40、80、160μmol/L黄芩素组对SGC996细胞转移抑制率与对照组比较,差异有统计学意义[(25.6±6.6)%、(57.3±7.9)%、(84.1±11.9)%比0,P0.05]。Western blot结果显示,黄芩素对ZFX有显著的抑制作用。结论黄芩素有良好的抗胆囊癌生物活性,其抗肿瘤效应的机制可能与下调ZFX蛋白表达有关。     

ZFX蛋白在胃癌组织中的表达与病理和预后关系的研究

目的:探讨胃癌组织中X染色体连锁锌指蛋白(ZFX)的表达,评价其与患者临床病理特征及预后的关系。方法:选取2013年1月—2015年12月经手术切除的胃癌组织标本,并收集其癌旁组织标本作为对照组。采用qRT-PCR方法检测43例胃癌组织及其对应癌旁组织中ZFX mRNA的表达,免疫组织化学(IHC)方法进一步检测114例胃癌及其癌旁组织中ZFX蛋白的表达情况,观察并探讨其与临床病理参数之间的关系,采用Kaplan-Meier对胃癌患者生存情况进行分析。结果:qRT-PCR结果显示ZFX mRNA在胃癌组织中的表达异常升高,明显高于癌旁组织;进一步IHC结果表明,ZFX蛋白的表达位于细胞核,胃癌组织中的表达阳性率约为76.3%,而癌旁组织中的阳性率仅有19.3%,差异有统计学意义(P0.01)。进一步分析显示,胃癌组织中ZFX蛋白的表达与患者的年龄、性别、肿瘤部位及TNM分期无关(P0.05),而与肿瘤分化程度、淋巴结转移、肿瘤的浸润深度密切相关(P0.05)。Kaplan-Meier生存曲线显示ZFX蛋白阴性表达组的总生存期高于阳性表达组(P0.05)。结论:ZFX在胃癌组织中于基因水平和蛋白水平均呈现异常高表达,并与患者临床病理参数和预后密切相关,提示ZFX参与胃癌的发生发展,有望成为胃癌潜在的预后指标。     

Zinc-finger protein X-linked is a novel predictor of prognosis in patients with colorectal cancer

Zinc-finger protein X-linked (ZFX) has been demonstrated to play an important role in the development of human malignancies. However, its prognostic significance in cancer patients remains unclear and less is known about its role in colorectal cancer (CRC). In this study, we found that the expression of ZFX in CRC tissues was significantly higher than that in corresponding normal tissues by quantitative real-time polymerase chain reaction and Western blot. Using immunohistochemistry, we explored the associations between protein expression of ZFX and clinicopathological parameters in 120 CRC cases. The results showed that ZFX expression was significantly associated with tumor differentiation (P = 0.022), tumor size (P = 0.037), tumor invasion (P = 0.027), lymph node metastasis (P = 0.042), distant metastasis (P = 0.011), and Dukes’ classification (P = 0.028). Moreover, according to Kaplan-Meier model, patients with high expression of ZFX had a significantly poorer prognosis compared to those with low expression of ZFX. Multivariate analysis suggested that high expression of ZFX was an independent prognostic factor for CRC patients. In conclusion, our findings for the first time demonstrated that ZFX expression may be associated with the progress of CRC and suggested that ZFX has the potential value to be an effective prognostic predictor for CRC patients.