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Dihydrotanshinone I potentiates the anti-tumor activity of cisplatin by activating ROS-mediated ER stress through targeting HSPD1 in lung cancer cells

Authors: Haizhen Lin,Hehuan Sui,Ying Yu,Chenjun Xie,Yiwei Shen,Liyuan Cheng,Jiaying Wang,Yun Yu,Congying Xie,Ri Cui
Publisher: Elsevier BV
Publish date: 2025-5
ISSN: 0014-2999 DOI: 10.1016/j.ejphar.2025.177378
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As a researcher exploring ROS-mediated cancer therapies, I am interested in replicating some aspects of your methodology, but I have a few questions regarding your experimental approach:

1. The study mentions using DCFH-DA staining to assess ROS levels after DHTS treatment. However, were any additional oxidative stress markers, such as lipid peroxidation assays or glutathione depletion measurements, considered to confirm ROS-mediated cytotoxicity? Would time-course experiments provide more insights into ROS accumulation dynamics?
2. The knockdown of HSPD1 was shown to enhance DHTS-induced apoptosis. Were multiple siRNA sequences tested to ensure target specificity, and was the silencing efficiency validated at both the mRNA and protein levels over different time points?
3. The CompuSyn software was used to determine the synergy between DHTS and cisplatin. Could you clarify whether the drug interaction was evaluated across different dose ratios, or was a fixed-ratio approach used? Additionally, how was the optimal dose combination selected for in vivo validation?
4. The study associates increased ATF4 and CHOP expression with ER stress-induced apoptosis. Were any functional inhibitors of ER stress (e.g., 4-PBA) used to confirm that apoptosis was directly mediated by ER stress rather than a secondary effect of ROS accumulation?
5. The xenograft model showed promising results for DHTS-cisplatin combination therapy. Were there any systemic toxicity evaluations (e.g., liver enzyme analysis, hematological parameters) to confirm the safety profile of this combination in comparison to cisplatin alone?

I would greatly appreciate your insights on these questions, as they would help me refine my approach in investigating ROS-based combination therapies in cancer research. Thank you for your time!

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