The role of EGFR in the pathogenesis of vaginal candidiasis

Abstract

Abstract:

Objective To investigate the role and molecular mechanism of EGFR in the pathogenesis of vaginal candidiasis. Methods Vaginal epithelial cells (VK2/E6E7 cells) were cultured. After stimulation with Candida albicans, the expression of immune factors in EGFR and possible related immune pathway factors were detected by real-time PCR. The EGFR-siRNA VK2/E6E7 cell model was constructed and co-cultured with Candida albicans. The cytokines secreted by vaginal epithelial cells and the protective ability against Candida albicans infection were detected by ELLISA and automatic cell detector. A mouse model of vaginal Candida infection was constructed. qPCR was used to detect the expression of EGFR and immune pathway factors in vaginal tissues. The changes of fungal load and inflammatory cells in vaginal tissues were detected after EGFR phosphorylation inhibitor blocking pathway. Results qPCR assay showed that EGFR, STAT3, GM-CSF and IL-1β were up-regulated in VK2/E6E7 cells infected with Candida albicans, and were statistically significant (P <0.05). ELLISA assay showed the expressions of GM-CSF, IL-8, IL-1β and MIP-3α were significantly decreased in EGFR-siRNA VK2/E6E7 cells infected with Candida albicans (P<0.05). The results of automatic cell detector showed that the defense capacity and cell activity of EGFR knockout cells after 30 hours' infection of Candida albicans were significantly lower than normal cells. The results of qPCR showed that the expression of EGFR, HER2, STAT3 and IL-8 following the vaginal infection of Candida albicans in the mouse was statistically significant (P<0.05); after local application of phosphorylation inhibitor, the lavage fluid load in the mouse vagina was significantly higher than that of the control group with EGFR pathway being blocked and there was a statistical difference on the 7th day after infection (P<0.05). Conclusion The EGFR and its pathway factors should play an important role in the pathogenesis of vaginal candidiasis.

Key words: EGFR, Candida albicans, vaginal candidiasis

CLC Number:

R519.3

ZHANG Jing-yun, PENG Jing-wen, WANG Qiong, GAO Ying, CHEN Wan-xin, SHEN Yong-nian, LI Dong-mei, SHE Xiao-dong, LIU Wei-da. The role of EGFR in the pathogenesis of vaginal candidiasis[J]. Chinese Journal of Mycology, 2020, 15(2): 65-71.

References

[1] Sobel JD. Recurrent vulvovaginal candidiasis[J].Am J Obstet Gynecol, 2016,214(1):15-21.
[2] Cassone A. Vulvovaginal Candida albicans infections:pathogenesis, immunity and vaccine prospects[J]. BJOG, 2015,122(6):785-794.
[3] 佘晓东, 沈永年, 韩峻松, 等. 用Oligo芯片和小鼠模型研究外阴阴道念珠菌病的发病机制[J].中华皮肤科杂志, 2009, 42:681-684.
[4] Sigismund S, Avanzato D, Lanzetti L. Emerging functions of the EGFR in cancer[J]. Mol Oncol,2018,12(1):3-20.
[5] Mascia F, Cataisson C, Lee TC, et al. EGFR regulates the expression of keratinocyte-derived granulocyte/macrophage colony-stimulating factor in vitro and in vivo[J]. J Invest Dermatol,2010,130(3):682-693.
[6] Shu DY, Wojciechowski M, Lovicu FJ. ERK1/2-mediated EGFR-signaling is required for TGFβ-induced lens epithelial-mesenchymal transition[J]. Exp Eye Res, 2019,178(1):108-121.
[7] Ray K, Ujvari B, Ramana V, et al. Cross-talk between EGFR and IL-6 drives oncogenic signaling and offers therapeutic opportunities in cancer[J]. Cytokine Growth Factor Rev, 2018,41(1):18-27.
[8] Chattopadhyay S, Veleeparambil M, Poddar D, et al. EGFR kinase activity is required for TLR4 signaling and the septic shock response[J]. EMBO Rep, 2015,16(11):1535-1547.
[9] Veleeparambil M, Poddar D, Abdulkhalek S, et al.Constitutively bound EGFR-mediated tyrosine phosphorylation of TLR9 is required for its ability to signal[J]. J Immunol,2018,200(8):2809-2818.
[10] Zhu W, Phan QT, Boontheung P, Solis NV, et al. EGFR and HER2 receptor kinase signaling mediate epithelial cell invasion by Candida albicans during oropharyngeal infection[J]. Proc Natl Acad Sci U S A,2012, 109:14194-14199.
[11] Wang SH, Wang SC, Chen PC, et al. Induction of cyclooxygenase-2 gene by Candida albicans through EGFR, ERK, and p38 pathways in human urinary epithelium[J]. Med Mycol,2017,55(3):314-322.
[12] Lionakis MS, Lim JK, Lee CC, et al. Organ-specific innate immune responses in a mouse model of invasive candidiasis[J]. J Innate Immun, 2011, 3(2):180-199.

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