Laboratory of Molecular Biology, East Biomedical Research Center, Mexican Institute of Social Security, Metepec 74360, Mexico.
Research Center of Microbiological Sciences, Institute of Sciences, Meritorious Autonomous University of Puebla, Puebla 72592, Mexico.
Mol Med Rep. 2020 Dec;22(6):5444-5453. doi: 10.3892/mmr.2020.11630. Epub 2020 Oct 23.
The altered expression of glycan antigens has been reported during cervix transformation, demonstrating increased mRNA levels of certain glycogenes. Human papillomavirus (HPV) is the aetiological agent of cervical cancer. High risk HPV E5 is considered an oncogene and has been implicated in cell transformation. E6 and E7 HPV oncoproteins modify the expression of certain glycogenes. The role of the E5 HPV protein in glycogene expression changes has not yet been reported. The aim of the present study was to determine the effects of HPV16 E5 oncoprotein on glycogene expression. For these, a microarray assay was performed using the HaCaT cell line and altered glycogenes were identified. The mRNA levels of certain glycogenes were determined via reverse transcription‑quantitative PCR (RT‑qPCR). Using in silico analysis, the present study identified that glycosylation pathways were altered by E5. Microarray analysis revealed alterations in certain glycogenes, including the upregulation of ST6GAL1, ST3GAL3, CHST2 and MANBA, and the downregulation of UGT2B15, GALNT11, NDST2 and UGT1A10. Increased mRNA levels were confirmed via RT‑qPCR for sialyltransferases genes. Additionally, in silico analysis was performed to identify glycosylation networks altered in the presence of the E5 oncoprotein. The analysis revealed that E5 could modify glycan sialylation, the N‑glycosylation pathway, keratan sulfate and glycosaminoglycan synthesis. To the best of our knowledge, the current study was the first to determine the role of the HPV16 E5 oncoprotein in glycogene expression changes. The results indicated that increased sialyltransferase mRNA levels reported in pre‑malignant and malignant cervical tissues could be the result of E5 oncoprotein expression. The results provide a possible role of HPV infection on glycosylation changes reported during cervix transformation.
糖抗原的表达改变已在宫颈转化过程中被报道,表明某些糖基因的 mRNA 水平增加。人乳头瘤病毒(HPV)是宫颈癌的病因。高危型 HPV E5 被认为是一种癌基因,并与细胞转化有关。E6 和 E7 HPV 癌蛋白改变某些糖基因的表达。HPV E5 蛋白在糖基因表达改变中的作用尚未被报道。本研究旨在确定 HPV16 E5 癌蛋白对糖基因表达的影响。为此,使用 HaCaT 细胞系进行了微阵列分析,并鉴定了改变的糖基因。通过逆转录-定量 PCR(RT-qPCR)测定某些糖基因的 mRNA 水平。通过计算机分析,本研究发现 E5 改变了糖基化途径。微阵列分析显示某些糖基因发生改变,包括 ST6GAL1、ST3GAL3、CHST2 和 MANBA 的上调,以及 UGT2B15、GALNT11、NDST2 和 UGT1A10 的下调。通过 RT-qPCR 对唾液酸转移酶基因的 mRNA 水平进行了验证。此外,还进行了计算机分析以鉴定存在 E5 癌蛋白时改变的糖基化网络。分析表明,E5 可以修饰聚糖唾液酸化、N-糖基化途径、硫酸角质素和糖胺聚糖合成。据我们所知,本研究首次确定了 HPV16 E5 癌蛋白在糖基因表达改变中的作用。研究结果表明,在癌前和恶性宫颈组织中报道的增加的唾液酸转移酶 mRNA 水平可能是 E5 癌蛋白表达的结果。这些结果为 HPV 感染在宫颈转化过程中报道的糖基化改变提供了一个可能的作用。