Debasis Nayak

Figure S8. Tumors in athymic nude mice orthotopically implanted with vehicle control and RGS11-OE MIA PaCa-2 cells.

Supplementary Materials & Methods

Figure S5. Chemosensitivity of PANC-1 RGS11-OE cells to chemotherapeutic drugs.

Figure S3. IHC staining for RGS11.

Table S12. Apical junction genes (top 50) positively enriched in RGS11-OE.

Table S2. Gene sets enriched in RGS11-OE MIA PaCa-2 cells (n=3).

Figure S7. Gene set enrichment analysis (GSEA) of RGS11-OE MIA PaCa-2 cells.

Table S7. KRAS signaling downstream pathway top 50 genes negatively enriched in RGS11-OE.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy associated with early metastasis, drug resistance, and poor outcomes. We previously demonstrated a putative tumor-suppressive role for concentrative nucleoside transporter 1 (CNT1) in PDAC. In this study, we demonstrate the regulator of G protein signaling (RGS) 11 as a key target of CNT1, with potent tumor-suppressive properties in PDAC. Compared with normal human pancreas, RGS11 expression is diminished in human PDAC tissues which correspond with the reduced patient survival times. In addition, quasimesenchymal pancreatic tumor cell lines with accelerated growth, metastatic propensity, and innate resistance to nucleoside analogs showed relatively lower RGS11 expression than their epithelial counterparts. Interestingly, RGS11 levels reversibly modulated the epithelial–mesenchymal transition of human PDAC cell lines influencing the chemotherapeutic sensitivities of anti-PDAC drugs. Additionally, stable lentiviral-mediated RGS11 expression reduced the cellular proliferation and colony establishment, increased the apoptotic index, and decreased the migratory and invasive abilities in quasimesenchymal tumor cell lines, whereas RGS11 depletion in epithelial tumor cell lines showed opposite effects. Global transcriptomic analysis revealed RGS11 replenishment in PDAC cells to suppress CD44-directed stemness features with significant reprogramming of the PDAC oncogenic landscape. Furthermore, RGS11 reduced the primary tumor burden and metastatic occurrence in a mouse model of PDAC. Together, these findings uncover RGS11 as a key target of CNT1 that exhibits therapeutic potential for intervention of aggressive PDAC.Implications:RGS11 identified as a downstream target of a gemcitabine transporter CNT1 exerts potent antitumorigenic features in PDAC with therapeutic and prognostic values.

Figure S2. Generation of CNT1 OE and KD clones in L3.6pl and evaluation of RGS11 protein expression.