Mechanism for Regulation of Melanoma Cell Death via Activation of Thermo-TRPV4 and TRPV2

Abstract

Background. Thermo-TRPs (temperature-sensitive transient receptor potential channels) belong to the TRP (transient receptor potential) channel superfamily. Emerging evidence implied that thermo-TRPs have been involved in regulation of cell fate in certain tumors. However, their distribution profiles and roles in melanoma remain incompletely understood. Methods. Western blot and digital PCR approaches were performed to identify the distribution profiles of six thermo-TRPs. MTT assessment was employed to detect cell viability. Flow cytometry was applied to test cell cycle and apoptosis. Calcium imaging was used to determine the function of channels. Five cell lines, including one normal human primary epidermal melanocytes and two human malignant melanoma (A375, G361) and two human metastatic melanoma (A2058, SK-MEL-3) cell lines, were chosen for this research. Results. In the present study, six thermo-TRPs including TRPV1/2/3/4, TRPA1, and TRPM8 were examined in human primary melanocytes and melanoma cells. We found that TRPV2/4, TRPA1, and TRPM8 exhibited ectopic distribution both in melanocytes and melanoma cells. Moreover, activation of TRPV2 and TRPV4 could lead to the decline of cell viability for melanoma A2058 and A375 cells. Subsequently, activation of TRPV2 by 2-APB (IC50 = 150 μM) induced cell necrosis in A2058 cells, while activation of TRPV4 by GSK1016790A (IC50 = 10 nM) enhanced apoptosis of A375 cells. Furthermore, TRPV4 mediated cell apoptosis of melanoma via phosphorylation of AKT and was involved in calcium regulation. Conclusion. Overall, our studies revealed that TRPV4 and TRPV2 mediated melanoma cell death via channel activation and characterized the mechanism of functional TRPV4 ion channel in regulating AKT pathway driven antitumor process. Thus, they may serve as potential biomarkers for the prognosis and are targeted for the therapeutic use in human melanoma.

Identification of differentially expressed methylated genes in melanoma versus nevi using bioinformatics methods

“…DTL is involved with the regulation of cell cycles and playing a role in the regulation of DNA damage, so we suggest hat it regulate cycle of melanocytes so that to regulate its progression. TRPV2 is an ion channel, it's activation mediated melanoma cell death (Zheng et al, 2019), which may play a negative role in melanoma development. PLOD3 was found to be highly expressed in lung cancer and gliomas (Tsai et al, 2018), which is associated with poor prognosis of them.…”

Epidermal TRPV4 ion channels regulate UVB induced sunburn by triggering inflammmasome activation and ERK signaling

“…These previous studies showed that TRPV4 activation caused severe cellular disarrangement, necrosis and apoptosis in melanoma. The experiments suggested that TRPV4 channel activation contributed to melanoma cell death via Ca ++ -influx, which also activated the AKT pathway with net anti-proliferative effects (49). One possible translational application is to enhance melanoma cell death via use of targeted UVB and/or topical TRPV4 agonists in conjunction with currently established melanoma treatments.…”

Peer Review #2 of "Identification of differentially expressed methylated genes in melanoma versus nevi using bioinformatics methods (v0.3)"

“…DTL is involved with the regulation of cell cycles and playing a role in the regulation of DNA damage, so we suggest hat it regulate cycle of melanocytes so that to regulate its progression. TRPV2 is an ion channel, it's activation mediated melanoma cell death [23] , which may play a negative role in melanoma development. PLOD3 was found to be highly expressed in lung cancer and gliomas [24] , which is associated with poor prognosis of them.…”