Table 4.

In Vitro Studies Demonstrating the Anticancer Effects of SGLT2 Inhibitors

First Author (Year)	Cancer Type	Cell Type	SGLT2i Treatment	Key Finding	Results and Proposed Mechanism
Villani et al (2016)24	Prostate, lung, liver, breast cancer	Prostate (PC3, 22RV-1), lung (A549, H1299), liver (HepG2), and breast (MCF7) cancer cells	CANA DAPA (0-100 μM)	Only CANA at clinically relevant concentrations ↓ proliferation and clonogenic survival of cancer cells alone and in combination with cytotoxic therapies	CANA: ↓ Glucose uptake, mitochondrial complex I–supported respiration ↓ ATP and lipogenesis ↑ Phosphorylation of AMPK
Li et al (2017)136	Resistant NSCLC	HCC827, H1975 carrying specific EGFR mutations	CANA (0-100 μM)	↓ Growth of NSCLC cell lines	↑ Apoptosis ↓ EGFR phosphorylation ↓ Phosphorylation of Akt and ERK
Kuang et al (2017)141	RCC	ACHN, A498, and CaKi-1	DAPA (0-4 μM)	↓ Cell growth in a dose- and time-dependent manner	↑ G1 phase arrest ↑ Apoptosis ↓ SGLT2 expression and glucose uptake
Kaji et al (2018)129	HCC	Huh7 and HepG2 cells	CANA (10 μM)	↓ Liver cancer cell growth and angiogenic activity	↓ Glycolytic metabolism ↑ G2/M arrest and apoptosis ↓ Phosphorylation of ERK, p38, and AKT
Hung et al (2019)130	HCC	Huh-7 and Hep3B	CANA (0-50 μM)	↓ Growth of HCC cells	↓ Expression of β-catenin ↑ Proteasomal degradation of β-catenin protein by ↑ phosphorylation of β-catenin Direct inhibition of PP2A activity
Nasiri et al (2019)23	Breast and colon cancer	E0771 breast tumors and MC38 colon tumors	CANA DAPA (5 μM and 5 mM)	↓ Cancer cell growth rate	CANA ↓ both glucose uptake and oxidation at clinically relevant concentrations (5 μM) DAPA did not alter glucose uptake or tumor cell division at clinically relevant concentrations (0.5 μM) but did at a suprapharmacologic concentration (5 mM)
Nakano et al (2020)25	HCC	Huh7 and HepG2	CANA (3 μM, 10 μM)	↓ Proliferation of HCC cells	Regulating metabolic reprogramming (alterations in metabolism of mitochondrial oxidative phosphorylation, fatty acid, and purine and pyrimidine) ↓ ATP synthase F1 subunit α Altered phosphorylation of AMPK
Zhou et al (2020)143	Breast cancer	MCF-7, ZR-75-1	CANA DAPA (0, 3.3, 11, 33, 100, 300 μM)	Both ↓ human breast cancer cells proliferation and growth	↑ Phosphorylation of AMPK ↓ Phosphorylation of p70S6K Cell cycle arrest in G1/G0 phase ↑ Cell apoptosis (AMPK-mediated cell cycle arrest and apoptosis)
Xu et al (2020)133	Pancreatic cancer	Capan-1 and PANC-1	CANA (20, 40, 60 and 80 μM)	↓ Pancreatic cancer cell proliferation and colony formation	↑ Apoptosis ↓ Glycolysis via the PI3K/AKT/mTOR pathway
Xie et al (2020)140	Cervical cancer	HeLa and C33A	EMPA (50 μM)	↓ Migration of cervical cancer cells and ↑ apoptosis	↑ AMPK/FOXA1 pathway and ↓ expression of Shh
Komatsu et al (2020)144	Breast cancer	MCF-7 cells	IPRA (1–50 μM)	↓ Breast cancer cell proliferation	SGLT2 inhibition-dependent hyperpolarization of MCF-7 cell membrane Mitochondrial membrane instability ↓ DNA synthesis at high dose
Papadopoli et al (2021)145	Breast cancer	MCF7, SKBR3 and BT-474, NT2197	CANA DAPA (25, 50 μM)	CANA ↓ proliferation of breast cancer cell lines DAPA showed modest effect	Antiproliferative effects were not affected by glucose availability or the level of expression of SGLT2 ↓ Mitochondrial respiration and total ATP production ↓ Glutamine metabolism
Ren et al (2021)134	Pancreatic cancer	PANC-1 and BxPC-3	CANA (1 μM) SOTA (3 nM)	↓ Proliferation and invasion of pancreatic cancer cells	↓ Hippo pathway ↓ YAP1 expression
Yamamoto et al (2021)137	Lung cancer	A549, H1975, and H520	CANA (1-50 μM)	↓ Growth of cells in a dose-dependent manner	↓ DNA synthesis ↓ S phase entry (induced G1 arrest) ↓ ERK and MAPK phosphorylation Did not induce apoptosis Tumor weight was not decreased by CANA in vivo
Wu et al (2022)22	Osteosarcoma	MNNG/HOS and MG-63	CANA (0.5, 1, or 2 μM)	↓ Osteosarcoma progression	Inducing immune cell infiltration ↑ STING/IRF3/IFN-β pathway ↓ AKT pathway
Wang et al (2022)142	Papillary thyroid cancer	TPC-1 and BCPAP	CANA (10 μM) DAPA (0, 20, 40, 80 μM)	↓ Thyroid cancer cells growth in a dose- and time-dependent manner DAPA ↓ proliferation of the same cells	(Mechanisms only done with CANA) Dependent on SGLT2 expression ↓ Invasion of thyroid cancer cell ↓ Glucose uptake ↓ glycolysis ↑ AMPK pathway ↓ AKT/mTOR pathway ↑ Cell cycle arrest at G1/S checkpoint ↑ DNA damage and ATM/CHK2 pathway activation ↑ Apoptosis
Shoda et al (2023)132	Glioblastoma	U251MG (human), U87MG (human), and GL261 (murine)	CANA (40 μM)	↓ Glioblastoma cell proliferation	Dependent on SGLT2 expression ↓ Glucose uptake ↑ AMPK phosphorylation ↓ p70S6K phosphorylation
Ding et al (2023)139	NSCLC, ovarian, pancreatic cancers	H292, SKOV3, MIA PaCa-2, primary NSCLC, ovarian and pancreatic cancer patient–derived cancer cells	CANA (20 μM)	SGLT2 is a positive regulator of PD-L1 (the interaction between SGLT2 and PD-L1 on the cell membrane is required for maintaining PD-L1 protein)	↓ PD-L1 protein expression ↑ Proteasomal degradation of PD-L1
Biziotis et al (2023)148	Human adenocarcinoma, squamous cell, NSCLC	Human adenocarcinoma (A549, H1299 and H1975), squamous cell (SK-MES-1), and large cell (H460) NSCLC cells	CANA (5-30 μM)	↓ Proliferation of all cell lines ↓ Clonogenic potential of A549, H1299, and H1975 cells	↓ Oxygen consumption rate ↑ Extracellular acidification rate ↑ AMPK activity ↓ mTOR activity ↓ (MAPK) ERK1/2 ↓ HIF-1α ↓ HDAC2

ATP = adenosine triphosphate; EGFR = epidermal growth factor receptor; FOXA1 = forkhead box A1; HCC = hepatocellular carcinoma; HDAC2 = histone deacetylase 2; HIF-1α = hypoxia-inducible factor-1α; IFN = interferon; IPRA = ipragliflozin; IRF3 = interferon regulatory factor 3; MAPK = mitogen-activated protein kinase; NSCLC = non-small-cell lung cancer; PD-L1 = programmed cell death-ligand 1; PP2A = protein phosphatase 2A; Shh = sonic hedgehog signaling molecule; SOTA = sotagliflozin; STING = stimulator of interferon genes; YAP1 = YES-associated protein 1; other abbreviations as in Tables 1 and 2.