In vitro investigation of the effect of paclitaxel, A chemotherapy agent, on cancer cachexia

Cachexia, is a multifactorial syndrome characterized by loss of skeletal muscle and adipose tissue, leading to progressive functional disorders that cannot be completely reversed by conventional nutritional support. Zinc-α2-glycoprotein (ZAG), one of the potential biomarkers of cachexia and encoded by the AZGP1 (Alpha-2-Glycoprotein 1, Zinc-Binding) gene, has been identified as a lipid mobilizing factor in adipose tissue. Adipose tissue is formed as a result of the adipogenesis period in which preadipocytes transform into mature adipocytes with the effect of transcription factors such as PPARγ and C/EBPα. It has been reported that AZGP1 inhibits adipogenesis and lipid accumulation by suppressing these transcription factors. However, it has been reported that AZGP1 triggers the conversion of white adipocytes to brown adipocytes and may contribute to adipose tissue loss by increasing fat catabolism in cachexia. Chemotherapy may contribute directly or indirectly to cachexia by causing nutritional and metabolic status changes as well as causing specific treatment damage. Paclitaxel is a chemotherapeutic agent frequently used in the treatment of many types of cancer. In this study, it was aimed to investigate the effect of paclitaxel on cancer cachexia through lipid accumulation and cachexia-related AZGP1 gene expression. In the study, human preadipocyte cells were grown in commercially purchased preadipocyte growth medium at 37°C and 5% CO2. Proliferating cells were seeded in 12-well cell culture dishes at 105 cells/well, and 50 nM paclitaxel + differentiation medium was applied at 3-day intervals for 15 days, which is the completion of differentiation. The application was made as paclitaxel + differentiation medium on day 0, day 3, day 6, day 9 and day 12 depending on time. At the end of the 15th day, Oil Red O staining was performed to detect lipid accumulation in the cells and total RNA was isolated for gene expression analysis. The effect of time-dependent paclitaxel on AZGP1 gene expression during adipogenesis was analyzed by RT-PCR. As a result of the study, it was determined that paclitaxel given during the adipogenesis process suppressed differentiation and lipid accumulation in human preadipocytes (p<0.01). As a result of gene expression analysis, AZGP1; Day 0, day 6, day 9 and day 12 showed 0.42, 0.44, 0.5 and 0.36 times less expression, respectively, but there was no change in AZGP1 expression on the 3rd day (p>0.05). PPARγ gene expression was 0.03 (p<0.05), 0.65, and 0.9-fold lower on day 0, day 3, and day 6, respectively, but 2.29 (p=0.015) and 3.99 (p=0.05) at day 9 and 12, respectively. It was determined that 0.003) times more expression. As a result, it was determined that paclitaxel may contribute to adipose tissue loss in cancer cachexia by suppressing lipid accumulation, but the change in cachexia-related AZGP1 and adipogenesis-related PPARγ gene expression levels in this process should be analyzed with further studies.