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. 2021 Oct 20;11:716246. doi: 10.3389/fonc.2021.716246

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Copyright © 2021 Wang, Liu, Liu, Cai, Yao, Chen, Song, Yuan, Tan, Hu, Xiang and Xiang

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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ST6GAL1 is a pivotal oncogenic mediator of the high-risk HPV 16 E6. (A) Analysis of ST6GAL1 expression in cervical scraping samples of five patient groups (n = 10 in each group). HPV-negative samples were used as control group, and ST6GAL1 expression values were normalized relative to expression of the housekeeping gene GAPDH by the 2^−ΔΔCt method. (B) The association of ST6GAL1 to E6 in E6-detected samples was analyzed by Spearman analysis (n = 30). (C) C33A and C33A-E6 (E6 stable expressing) cells were transiently transfected with short hairpin RNA targeting ST6GAL1 (shST6GAL1) or a non-targeting control (shNTC). Then, protein expression of E6 and ST6GAL1 were detected by Western blot. (D) Proliferation of indicated cells were detected by CCK-8 assay. (E) The clones of indicated cells were visualized by crystal violet staining. (F) Statistical analysis of colony numbers in C33A and C33A-E6 cells. (G, H) Apoptosis of indicated cells were detected by flow cytometry, and then, apoptotic rates were statistically analyzed. (I) Western blot was used to detect the expression of apoptosis-related proteins. (J, K) Migrative abilities of indicated cells were detected by wound-healing assay and statistically analyzed. (L, M) Invasive abilities of indicated cells were detected by Transwell assay and statistically analyzed. Data were presented as mean ± SD from the three independent replicates. *p < 0.05; **p < 0.01; ***p < 0.001.