SH-SY5Y cells are neuroblastoma cells that exhibit neuron-like behavior and rely more heavily on basal autophagy to maintain their homeostasis, and they are more sensitive to mitochondrial dysfunction. While HeLa cells are cervical cancer cells that are more resistant to mitochondrial dysfunction, leading to their proliferation compared to SH-SY5Y cells. Try using a lower concentration for SH-SY5Y cells to detect the cytotoxicity threshold. In addition, you can use different time points for HeLa cells (24, 48, 72) hrs to reveal the dose-dependent toxicity.

Nourhan Abdelmaksoud Thank you for the clarification.

Hello Gaurav,

Bafilomycin A1 is a drug that inhibits V-ATPase, a protein complex essential for lysosomal acidification. This inhibition disrupts autophagy by preventing the fusion of autophagosomes with lysosomes.

SH-SY5Y cells may be more sensitive to disruptions in autophagy as they may rely more heavily on autophagy for maintaining cellular homeostasis and survival, making them more susceptible to the effects of autophagy inhibition. So, SH-SY5Y cells might be more vulnerable to the toxic effects of accumulating cellular waste (proteins, damaged organelles) when autophagy is blocked by Bafilomycin A1. On the other hand, HeLa cells are generally more adaptable and may have a more robust autophagy pathway or a different reliance on autophagy for survival.

I suggest you try performing dose-response experiments with Bafilomycin A1 which can help determine the specific concentration at which cell death occurs and whether there is a threshold effect. Try out assays to assess autophagy flux, such as LC3 turnover and p62 degradation to obtain a comprehensive understanding of the autophagic process. 

Best,

SH-SY5Y, a human neuroblastoma cell line, is rather sensitive to autophagy inhibition in neuronal progenitor cells. The autophagy inhibition is actively researched as a therapeutic strategy for neuroblastoma patients. If you want to study autophagy inhibition in neuronal cells, you can either try to differentiate the SH-SY5Y with retinoic acid and BNDF or use primary neuronal cultures. You can also use chloroquine different approach to autophagy inhibition. Unless you are specifically studying the effects of Bafilomycin A1.