Deregulation of LIMD1-VHL-HIF-lα-VEGF pathway is associated with different stages of cervical cancer
Deregulation of LIMD1-VHL-HIF-lα-VEGF pathway is associated with different stages of cervical cancer
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DOI：
10.1042/bcj20170649
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发表时间：
2018-05-31
影响因子：
4.1
通讯作者：
Panda, Chinmay Kumar
中科院分区：
文献类型：
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作者：
Chakraborty, Chandraditya;Mitra, Sraboni;Panda, Chinmay Kumar
为了解正常宫颈上皮基底-副基底层及宫颈癌不同时期细胞应力的变化机制，我们分析了细胞应力在宫颈癌不同时期的变化，HIF-1 α及其相关基因LIMD 1、VHL和VEGF在无病正常宫颈（n = 9）、肿瘤的邻近正常宫颈（n = 70）、宫颈上皮内瘤变（CIN; n = 32）、宫颈癌（CACX; n = 174）样品和两种CACX细胞系。在正常宫颈上皮的基底-副基底层中，LIMD 1显示高蛋白表达，而VHL的低蛋白表达与HIF-Ia和VEGF的高表达一致，而与HPV-16（人乳头瘤病毒16）感染无关。这与正常子宫颈基底-副基底层LIMD 1启动子甲基化水平低和VHL水平高相一致。在宫颈癌的不同阶段，LIMD 1的表达明显降低，而VHL的表达无明显变化。这与其在肿瘤不同阶段的频繁甲基化一致。在宫颈癌的不同分期中，HIF-1 α和VEGF在基底-副基底层的表达均较高，且与LIMD 1和VHL的表达呈负相关。这通过在CACX细胞系中使用5-氮杂-2 '-脱氧胞苷的去甲基化实验来验证。CIN/CACX中LIMD 1 α和VHL的额外缺失通过降低基因表达在宫颈癌发生过程中提供了额外的生长优势，并与患者的不良预后相关。我们的数据表明，HIF-1 α及其靶基因VEGF在正常宫颈基底-副基底层的过度表达是由于VHL启动子甲基化导致的频繁失活。在宫颈癌的不同阶段，VHL和LIMD 1的额外甲基化/缺失保持了这种特征。
To understand the mechanism of cellular stress in basal-parabasal layers of normal cervical epithelium and during different stages of cervical carcinoma, we analyzed the alterations (expression/methylation/copy number variation/mutation) of HIF-1 alpha and its associated genes LIMD1, VHL and VEGF in disease-free normal cervix (n = 9) , adjacent normal cervix of tumors (n = 70), cervical intraepithelial neoplasia (CI N; n = 32), cancer of uterine cervix (CACX; n = 174) samples and two CACX cell lines. In basal-parabasal layers of normal cervical epithelium, LIMD1 showed high protein expression, while low protein expression of VHL was concordant with high expression of HIF-la and VEGF irrespective of HPV-16 (human papillomavirus 16) infection. This was in concordance with the low promoter methylation of LIMD1 and high in VHL in the basal-parabasal layers of normal cervix. LIMD1 expression was significantly reduced while VHL expression was unchanged during different stages of cervical carcinoma. This was in concordance with their frequent methylation during different stages of this tumor. In different stages of cervical carcinoma, the expression pattern of HIF-1 alpha and VEGF was high as seen in basal-parabasal layers and inversely correlated with the expression of LIMD1 and VHL. This was validated by demethylation experiments using 5-aza-2'-deoxycytidine in CACX cell lines. Additional deletion of LIMD1 alpha and VHL in CIN/CACX provided an additional growth advantage during cervical carcinogenesis through reduced expression of genes and associated with poor prognosis of patients. Our data showed that overexpression of HIF-1 alpha and its target gene VEGF in the basal-parabasal layers of normal cervix was due to frequent inactivation of VHL by its promoter methylation. This profile was maintained during different stages of cervical carcinoma with additional methylation/deletion of VHL and LIMD1.