GABRD is upregulated in breast cancer and predicts a poor prognosis.

A Breast cancer expression profile obtained from the TCGA database (blue) and the normal tissue RNA-seq profile obtained from the TCGA database and GTEx database (red). The expression levels of GABAAR and GABABR subunits were shown. GABRD were most significantly upregulated in breast cancer tissue than normal tissue. *p<0.05, **p<0.01, ***p<0.001; B Expression of GABRD in breast cancer tissues (n=150) and paracarcinoma tissues (n=52) detected by IHC; C GABRD IHC score between tumor tissue (tumor) and paracarcinoma tissue (normal). ***p<0.001; D Kaplan–Meier survival analysis of breast cancer patients with low and high GABRD expression levels, p=0.014.

Expression patterns of GABRD in breast cancer tissues and para-carcinoma tissues revealed in immunohistochemistry analysis, p<0.001.

Relationship between GABRD expression and tumor characteristics in patients with breast cancer. *p<0.05

GABRD knockdown inhibited breast cancer development in vitro.

A Cell count assay; B Wound healing assay; C Transwell assay; D Rate of apoptosis determined by flow cytometry; E Percentage of cells in each phase of the cell cycle determined by flow cytometry. *p<0.05, **p<0.01, ***p<0.001.

Knockdown of GABRD inhibits tumorigenesis and tumor growth of breast cancer in vivo.

A Photographs of tumors excised 59 days after the inoculation of breast cancer cells into nude mice; B Fluorescence intensity of tumors detected in animal imaging scanned and C quantified as a representation of the tumor burden; D Tumor weights were measured after the mice were sacrificed on Day 59; E Tumor volume changes with time after inoculation; the data are shown as the mean±SD (n≥3). * p < 0.05, ** p < 0.01, *** p < 0.001.

GABRD knockdown inhibits breast cancer by interacting with and regulating CDK1.

A Volcano plot and B heatmap showing the differentially expressed genes identified by gene microarray analysis of shGABRD and shCtrl cells. Several differentially expressed genes were selected for further verification by C qPCR and D western blotting; E Molecular interaction network involving GABRD and the selected differentially expressed genes constructed based on IPA; F Coimmunoprecipitation assay performed to verify the interaction between GABRD and CDK1.

GABRD-regulated tumor inhibition is CDK1 dependent.

A Cell counting assay; B Apoptosis analyzed by flow cytometry; C Wound healing assay; D Transwell assay; the data are shown as the mean±SD (n≥3). * p<0.05, ** p<0.01, *** p<0.001.

CDK1 overexpression reverses the inhibition of breast cancer induced by GABRD knockdown in an orthotopic mouse model.

A Photographs of tumors excised 42 days after the inoculation of breast cancer cells into nude mice; B Tumor weight measured after sacrifice; C Tumor volume changed with time after inoculation; the data are shown as the mean±SD (n≥3). * p < 0.05, ** p < 0.01, *** p < 0.001.