Supplementary MaterialsAdditional file 1: Table S2

Supplementary MaterialsAdditional file 1: Table S2. using western blot and q-RT-PCR. The data represent the means SD of three impartial experiments. (TIF 196 kb) 13046_2018_842_MOESM4_ESM.tif (196K) GUID:?E7371B61-3C5B-4F04-B5C7-E9FD34EECD1D Additional file 5: Figure S3. (a-b) Mel-RM with or without KLF5 or KLF8 knockdown were treated with 3?M TM. Cell lysates were then subjected to western blot analysis using the indicated antibodies. (TIF 192 kb) 13046_2018_842_MOESM5_ESM.tif (192K) GUID:?FABFBDE0-A819-49F7-BC5B-58CAD56833D1 Additional file 6: Figure S4. (a) the mRNA levels of VEGF were analysed by q-RT-PCR in KLF4 WT, KLF4 KO or KLF4 KO?+?Flag-KLF4 Mel-RM cells with or without 3?M TM treatment for the indicated occasions. The data represent the means SD of three impartial experiments. ** em p /em ? ?0.01, *** em p /em ? ?0.001 vs. control. (b-e) The expression levels of NUCB2 were detected by western blot and q-RT-PCR assays in Mel-RM cells Sulbutiamine with or without KLF5 or KLF8 knockdown under TM treatment. The data represent the means SD of three impartial experiments. *** em p /em ? ?0.001 vs. control. (TIF 252 GADD45B kb) 13046_2018_842_MOESM6_ESM.tif (252K) GUID:?F6E5F133-C372-43C2-BD29-D173F0A6F637 Additional file 7: Figure S5. NUCB2 was transfected in a stable manner into Mel-RM cells with or without KLF4 knockout. Cell lysates were then subjected to western blot analysis using the indicated antibodies. (TIF 69 kb) 13046_2018_842_MOESM7_ESM.tif (70K) GUID:?89FD83B2-D1E7-4954-8054-DBBDD0EB475F Abstract Background Adaptation to ER stress has been indicated to play Sulbutiamine an important role in resistance to therapy in human melanoma. However, the relationship between adaptation to ER stress and cell metastasis in human melanoma remains unclear. Methods The relationship of adaptation to ER stress and cell metastasis was investigated using transwell and mouse metastasis assays. The potential molecular mechanism of KLF4 in regulating the adaptation to ER stress and cell metastasis was investigated using RNA sequencing analysis, q-RT-PCR and western blot assays. The transcriptional regulation of nucleobindin 2 (NUCB2) by KLF4 was identified using bioinformatic analysis, luciferase assay, and chromatin immunoprecipitation (ChIP). The clinical significance of KLF4 and NUCB2 was based on human tissue microarray (TMA) analysis. Results Here, we exhibited that KLF4 was induced by ER stress in melanoma cells, and increased KLF4 inhibited cell apoptosis and promoted cell metastasis. Further mechanistic studies revealed that KLF4 directly bound to the promoter of NUCB2, facilitating its transcription. Additionally, an increase in KLF4 promoted melanoma ER stress resistance, tumour growth and cell metastasis by regulating NCUB2 expression in vitro and in vivo. Elevated KLF4 was found in human melanoma tissues, which was associated with NUCB2 expression. Conclusion Our data revealed that the promotion of ER stress resistance via the KLF4-NUCB2 axis is essential for melanoma cell metastasis, and KLF4 may be a promising specific target for melanoma therapy. Electronic supplementary material The online version of this article (10.1186/s13046-018-0842-z) contains supplementary material, which is available to authorized users. strong class=”kwd-title” Keywords: Melanoma, ER stress, Apoptosis, KLF4, Metastasis Background Melanoma is the most aggressive skin cancer and is associated with a high mortality rate [1]. In the past, multiple strategies have been used for melanoma treatment. For primary melanoma, surgical resection was the best option and the patients have a good post-treatment prognosis. For metastatic melanoma, only a few options were clinically available for treating the disease such as targeting BRAF and MEK using small molecule inhibitors, immunotherapeutic antibodies against the immune checkpoints T-lymphocyte-associated antigen 4 (CTLA-4) and programmed cell-death protein 1 (PD-1) and the altered oncolytic herpes virus talimogene laharparepvec (T-VEC) and the 5-12 months survival rate of patients has been improved to some extent [2C14]. However, similar to other cancers, distant metastasis is still a major obstacle to the treatment of melanoma. Thus, understanding the potential molecular mechanisms of tumour metastasis is usually important for improving the overall prognosis of patients with melanoma. Metastasis is usually a multi-step process releasing tumour cells from a primary lesion Sulbutiamine to a disparate organ or organs within the body. Tumour cells change their characteristics throughout the process, which enables them to proliferate and migrate, invading surrounding tissues [15]. When tumours metastasise, the cells undergone diverse microenvironments such.