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The HSF1/miR-135b-5p axis induces protective autophagy to promote oxaliplatin resistance through the MUL1/ULK1 pathway in colorectal cancer

Abstract

Oxaliplatin (oxa) is widely used in the treatment of colorectal cancer (CRC), but the development of oxaliplatin resistance is a major obstacle to the therapeutic efficacy in patients. MicroRNAs (miRNAs), endogenous noncoding RNAs measuring between 22 and 24 nucleotides, have been shown to be involved in the development of CRC drug resistance. However, the mechanism by which differentially expressed miRNAs induce chemotherapy resistance in CRC has not been fully elucidated to date. Here, we showed the differentially expressed miRNAs in oxaliplatin-sensitive and oxaliplatin-resistant CRC cells through miRNA microarray technology and found that miR-135b-5p was significantly increased in oxaliplatin-resistant cells. And miR-135b-5p was increased in the serum of colorectal cancer patients. More importantly, the miR-135b-5p level in the serum of oxaliplatin-resistant patients was further increased compared to that of oxaliplatin-sensitive patients. Recent studies have shown that protective autophagy is an important mechanism that promotes drug resistance in tumors. The potential role of miR-135b-5p in inducing protective autophagy and promoting oxaliplatin resistance was evaluated in two stable oxaliplatin-resistant CRC cell lines and their parental cells. We further identified MUL1 as a direct downstream target of miR-135b-5p and showed that MUL1 could degrade the key molecule of autophagy, ULK1, through ubiquitination. Mouse xenograft models were adopted to evaluate the correlation between miR-135b-5p and oxaliplatin-induced autophagy in vivo. Furthermore, we also investigated the regulatory factors for the upregulation of miR-135b-5p in CRC cells under oxaliplatin chemotoxicity. These results indicated that miR-135b-5p upregulation in colorectal cancer could induce protective autophagy through the MUL1/ULK1 signaling pathway and promote oxaliplatin resistance. Targeting miR-135b-5p may provide a new treatment strategy for reversing oxaliplatin resistance in CRC.

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Fig. 1: The increased expression of miR-135b-5p was associated with oxaliplatin resistance in colorectal cancer.
Fig. 2: miR-135b-5p promotes colorectal cancer proliferation and oxaliplatin resistance.
Fig. 3: Oxaliplatin induces protective autophagy in colorectal cancer.
Fig. 4: miR-135b-5p promotes oxaliplatin resistance in colorectal cancer cell lines by inducing protective autophagy.
Fig. 5: MUL1 is predicted and verified as the direct target of miR-135b-5p.
Fig. 6: The miR-135b-5p/MUL1 axis regulated autophagy activity by ubiquitinating ULK1.
Fig. 7: miR-135b-5p promotes oxaliplatin resistance by inducing protective autophagy in vivo.
Fig. 8: Inhibition of autophagy could reverse oxaliplatin resistance induced by miR-135b-5p in colorectal cancer.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (Nos. 82072664, 81772629, 81802363, 81702431, 81772843, 81974374) and the Demonstrative Research Platform of Clinical Evaluation Technology for New Anticancer Drugs (No. 2018ZX09201015). This work was also supported by the Tianjin Science Foundation (Nos. 18JCQNJC81900, 18JCYBJC92000, 18JCYBJC25400, 18JCYBJC92900, 20JCYBJC00100) and the Science & Technology Development Fund of the Tianjin Education Commission for Higher Education (2018KJ046, 2017KJ227). The funders had no role in the study design, the data collection and analysis, the interpretation of the data, the writing of the report, and the decision to submit this article for publication.

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HW, XW and HZ performed most of the experiments, analyzed the data, and wrote the manuscript. TD, RL, YL and HL reviewed and edited the manuscript. MB, TN, JW, GS performed some of the experiments. YB designed the experiments and is the guarantor of this work, had full access to all of the data in the study, takes responsibility for the integrity of the data and the accuracy of the data analysis.

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Correspondence to Yi Ba.

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Wang, H., Wang, X., Zhang, H. et al. The HSF1/miR-135b-5p axis induces protective autophagy to promote oxaliplatin resistance through the MUL1/ULK1 pathway in colorectal cancer. Oncogene 40, 4695–4708 (2021). https://doi.org/10.1038/s41388-021-01898-z

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