Chinese herbal compound SanHuang decoction reverses axitinib resistance in ccRCC through regulating immune cell infiltration by affecting ADAMTS18 expression.

This investigation aims to study the reversal effect of the Chinese herbal compound SanHuang decoction on axitinib resistance in clear cell renal cell carcinoma (ccRCC) cells and its mechanistic role by employing cellular and mouse models. Axitinib-resistant ccRCC cell lines (A498-DR and 786-O-DR) were cultured and treated with SanHuang decoction. The apoptosis and migration of tumor cells were observed by flow cytometry and wound healing assays, respectively, and the expression of a disintegrin-like and metalloprotease with thrombospondin type 1 motif 18 (ADAMTS18) was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting (WB). In addition, A498-DR cells were inoculated into mice to establish tumorigenic models, and the models were treated with normal saline, axitinib, or different concentrations of SanHuang decoction plus axitinib. Then, the tumor diameter in each group was measured, and the expression of ADAMTS18 was evaluated by RT-PCR, WB and immunohistochemistry. In addition, the distribution of T cells (CD45+, CD4+, CD8+) and PD-L1 expression was analyzed by flow cytometry to evaluate the level of immune cell infiltration. SanHuang decoction significantly reduced the proliferative activity of axitinib-resistant tumor cells and enhanced the sensitivity of tumors to axitinib in vitro (cell lines) and in mice. In the SanHuang decoction group, the expression level of ADAMTS18 was increased to some extent, and several phenomena were observed, including (1) subcutaneous transplanted tumors grew slower, (2) the CD45+/PD-L1 ratio was decreased and (3) the proportions of CD8+ and CD4+ T cells were increased. Overexpression of ADAMTS18 was synergistic with SanHuang decoction treatment to jointly improve tumor immune infiltration and inhibit immune escape. Pearson correlation analysis of sample data showed that there was a negative correlation between the expression of ADAMTS18 and PD-L1 in tumor tissues. In conclusion, the Chinese herbal compound SanHuang decoction can reverse axitinib resistance in ccRCC cells by regulating immune cell infiltration and affecting ADAMTS18 expression.