Glycolysis is essential for chemoresistance induced by transient receptor potential channel C5 in colorectal cancer

Abstract

BackgroundElevated intracellular Ca2+ ([Ca2+]i) level could lead to [Ca2+]i overload and promote apoptosis via different pathways. In our previously study, up-regulated expression of transient receptor potential canonical channel (TRPC5) was proven to increase [Ca2+]i level, and resulted in chemoresistance whereas not apoptosis in human colorectal cancer (CRC) cells. The ATP-dependent homeostatic maintenance of resting [Ca2+]i should be important in this process. Increased glycolysis was found to be an important adenosine triphosphate (ATP) source in cancer. This study aimed to explore the potential mechanism of aerobic glycolysis in transient receptor potential channel TRPC5 induced chemoresistance.MethodsIn this study, we examined glucose transporter 1 (GLUT1) expression, glucose consumption and celluar ATP production to determine glycolytic activity. Real-time PCR and western blot were analyzed to determine TRPC5 expression at the mRNA and protein levels in human CRC cells (HCT-8, LoVo), and fluorouracil (5-Fu) resistant CRC cells (HCT-8/5-Fu, LoVo/5-Fu). 3-bromopyruvate (3-BP) and 2-Deoxy-D-glucose (2DG) were used to inhibit glycolysis. Glycolytic activity, intracellular Ca2+ ([Ca2+]i) and the half maximal inhibitory concentration of 5-Fu (5-Fu IC50) were measured. Western blot was analyzed to determine cleaved Caspase-3 protein level. Flow cytometry was performed to detect the apoptosis rates. Immunohistochemistry staining was performed to determine TRPC5 and GLUT1 expression level in human CRC tissues.ResultsOverproduced of TRPC5 and increased glycolysis were found in HCT-8/5-Fu and LoVo/5-Fu than in HCT-8 and LoVo cells. Compared to HCT-8 cells, the HCT-8/5-Fu cells showed higher [Ca2+]i levels which decreased after treated with TRPC5-specific shRNA. Furthemore, inhibition of glycolysis resulted in decreased ATP production, elevation of [Ca2+]i level and cleaved caspase-3, increased apoptotic cells rate, and a remarkable reversal of 5-Fu resistance in HCT-8/5-Fu cells, while showed no effect in HCT-8 cells. BAPTA-AM, a [Ca2+]i chelator, could reduce the elevation of cleaved caspase-3 and increased apoptotic cells rate due to glycolysis inhibition. Advanced CRC patients with high expression of TRPC5/GLUT1 displayed poorer chemotherapy outcome, and notably, the significant association between high TRPC5 expression and chemoresistance is GLUT1 expression level dependent.ConclusionsWe demonstrated the essential role of glycolysis in TRPC5 induced chemoresistance in human CRC cells via maintaining [Ca2+]i homeostasis.

<p>MiR-138 Suppresses the PDK1 Expression to Decrease the Oxaliplatin Resistance of Colorectal Cancer</p>

“…Recently, studies report that chemoresistant cancer show high level of glycolysis. 24,25 However, the potential mechanism for illustrating the association between glycolysis and chemoresistance is still unclear. In the present study, we found that the oxaliplatin-resistant CRC cells consumed more amount of glucose and exhibited higher level of glycolysis compared to the routine CRC cells.…”

“…We next evaluated the difference of glycolysis between HT29/R (SW480/R) cells and their parental HT29 (SW480) cells, because some studies have reported that glycolysis is essential for chemoresistance in some cancers. 24,25 As shown in Figure 2A, HT29/R and SW480/R cells consumed more amount of glucose compared to the HT29 and SW480 cells. Furthermore, we observed that HT29/R and SW480/R cells produced more amount of lactate compared to the HT29 and SW480 cells ( Figure 2B).…”

Crab Ash Extract has Anti-Proliferative Effects on SK-MEL-28 Melanoma Cells and Induces a Cellular Stress Response and Metabolic Changes

“…43 Moreover, ENO1 knockdown was found to reduce glycolysis, increase OXPHOS, and cause growth arrest. 44,45 Other studies have found increased aerobic glycolysis is important in maintaining chemo-resistance, and it has been hypothesised that glycolytically-derived ATP contributes to, among other things, drug inactivation and increased intracellular survival signalling. 46 The upregulation of the mitochondrial MICOS complex subunit MIC60 (IMMT) and mitochondrial 60kDa HSP suggests there may have been some mitochondrial damage/stress associated with the treatment -MIC60 is a core unit of MICOS, which regulates cristae morphology and protein transport; 47 while mitochondrial HSP60 maintains proteins in an unfolded state to enable transport across the inner mitochondrial membrane.…”

Polymorphisms in Glycolysis-Related Genes Are Associated with Clinical Outcomes of Paclitaxel-Cisplatin Chemotherapy in Non-Small Cell Lung Cancer

“…Studies have revealed that enhanced glycolysis is associated with aggressiveness and poor prognosis in multiple cancers [16][17][18]. Accumulating data have also indicated that elevated aerobic glycolysis in cancer cells is critical for chemoresistance [18][19][20]. Taken together, the diverse roles of aerobic glycolysis in cancer suggest that alterations in the glycolytic pathway may influence chemotherapy outcomes.…”