Effect of Thapsigargin on ER stress and cisplatin resistance in cisplatin-resistant Calu1 and H2030 non-small cell lung cancer cell lines
Project no. 23794 · Status: Kapanmış · 2022
Summary
<p>Effect of thapsigargin on endoplasmic reticulum (ER) stress and cisplatin resistance in cisplatin-resistant Calu1 and H2030 NHDAK cells</p><p>Objective: Cancer cells may be naturally resistant to cisplatin, an anticancer drug used in the treatment of solid tumors, or they may develop resistance during chemotherapy. The SERCA inhibitor thapsigargin triggers ER stress and stress-induced apoptosis. Our study aimed to investigate the potential of thapsigargin to prevent cisplatin resistance in cisplatin-resistant NHDAK cells.</p><p>Method: MTT was performed to determine the cytotoxic effects of cisplatin and thapsigargin in Calu1 and H2030 NHDAK cells and their cisplatin-resistant strains (SD-Calu1 and SD-H2030), qRT-PCR to determine mRNA levels of ER stress genes, Annexin V/PI staining for apoptosis, wound healing and colony formation assays for migration.</p><p>Results: SD-Calu1 cells developed resistance to cisplatin 7.5 times compared to Calu1 cells; SD-H2030 cells developed resistance 3.13 times compared to H2030 cells. Cisplatin resistance was significantly reduced in SD-Calu1 and SD-H2030 cells treated with thapsigargin. In Calu1 cells treated with thapsigargin, GRP78 expression increased 14.31 times and XBP1 expression increased 2.98 times. In SD-Calu1 cells, GRP78 expression increased 5.5 times, PERK expression increased 9.6 times, XBP1 expression increased 3.4 times and ATF4 expression increased 6.6 times. In H2030 cells, GRP78 expression increased 4.4 times after thapsigargin application. In SD-H2030 cells, GRP78 expression started to increase earlier and remained high for a longer period of time. In H2030 cells, XBP1 expression increased 3.3 times and ATF4 expression increased 4.2 times significantly. In SD-H2030 cells, XBP1 expression increased 8-fold and ATF4 expression increased 4.2-fold.</p><p>Thapsigargin increased apoptosis in Calu1, SD-Calu1 and H2030 cells. It was determined that colony formation efficiency decreased by 60.27% for Calu1; 89.64% for SD-Calu1; 51.46% for H2030 and 55.96% for SD-H2030. While wound healing areas closed in 48 hours in Calu1, H2030 and SD-H2030 cells, they closed by 33.3%, 14.77% and 20.94% in thapsigargin-treated cells, respectively. While wound healing areas closed in 72 hours in SD-Calu1 cells, this rate was determined as 20.82% in thapsigargin-treated cells.</p><p>Conclusion: The findings indicate that thapsigargin has the potential to be an effective agent in reversing cisplatin resistance in NSCLC.</p>