Integrated analysis and functional validation reveal KCNQ1 tumor suppressor targeting by dahuang Zhechong Pills via cuproptosis modulation in colorectal cancer.

[BACKGROUND] Colorectal cancer(CRC) makes difficulties to human beings. KCNQ1 is a possible tumor suppressor among potassium channels, but it is not yet known if there is any tumor suppression and whether it is epigenetically regulated in CRC. Dahuang Zhechong Pills (DHZCP) is a traditional Chinese medicine with anti-tumor effects, but its mechanisms, especially the KCNQ1 and cuproptosis pathways, still need to be elucidated. The motivation of this study is to draw up a high-resolution mechanistic map of Dahuang Zhechong Pills (DHZCP) by combining genetic causality with function validation, so as to raise both the academic value and clinical value of traditional Chinese medicine (TCM) in the management of CRC.

[METHODS] The integrative causal-validation framework was carried out. This study used the SMR based on the summary data to measure the causal effect of methylation on KCNQ1 expression, network pharmacology and molecular docking were used to predict the DHZCP targets, and in vitro studies were conducted with HCT-116 CRC cells. In vitro study was carried out in KCNQ1 overexpression (KCNQ1-OE) and DHZCP treated cells to study the effect of cell proliferation, apoptosis, migration, invasion, oxidative stress, and intracellular copper and expression of cuproptosis related protein (FDX1, DLAT, LIAS). Set up of clinical potency by systematically meta-analytical researches.

[RESULTS] SMR showed that KCNQ1 methylation negative regulate KCNQ1. KCNQ1 overexpression inhibited HCT-116 cell proliferation, migration, invasion and induced apoptosis, Oxidative stress. DHZCP-containing serum replicating and increasing these ones. Both decreased FDX1, DLAT and LIAS, increased ROS, MDA, 4-HNE and intracellular copper. DHZCP components bound directly to KCNQ1 in silico and multi-target actions against CRC were implied by network pharmacology. Meta-analysis of the clinical benefits of DHZCP in cancer therapy.

[CONCLUSION] KCNQ1 is a tumor suppressor of CRC that is DNA methylated. DHZCP in combination with KCNQ1 overexpression exhibits anti-CRC effects through the regulation of cuproptosis-related pathways, cuproptosis is promoted, oxidative stress is enhanced, and copper accumulates, thus supporting the clinical application prospects of DHZCP in CRC.