Regulatory Mechanisms of Cell Death Induced by Lactobacillus buchneri, Chlorella vulgaris Probiotics, and Chitosan-Conjugated Phytochemicals in Pancreatic Cancer Cells

Background and Aim

This study explores the therapeutic potential of natural compounds on pancreatic cancer cell lines (Panc-1). The main objective is to assess the effects of Lactobacillus buchneri supernatant, chitosan nanoparticles conjugated with Genistein (GCNPs), chitosan nanoparticles containing Epigallocatechin-3-Gallate (EGCG) and Chlorella vulgaris. Pancreatic cancer, especially Pancreatic Ductal Adenocarcinoma (PDAC), is highly aggressive and resistant, with few treatment options and low survival rates due to late diagnosis. The study aims to investigate the combined effects of these compounds on inducing apoptosis and altering gene expression related to cell death in pancreatic cancer cell lines. By evaluating these natural compounds, the research seeks to identify potential therapeutic agents for improving treatment outcomes in pancreatic cancer.

Materials and Methods

To evaluate the effects of treatments on Panc-1 cells, IC₅₀ values were determined using MTT assays at 24 and 48 hr. Cells were treated with Lactobacillus buchneri supernatant, chitosan nanoparticles conjugated with Genistein (GCNPs), chitosan nanoparticles containing EGCG and Chlorella vulgaris. Post-treatment, RNA was extracted for gene expression analysis via qPCR, focusing on CASP9, CYCLIND1, MMP2, VEGF, Bax, and Bcl-2. The expression levels were quantified to assess apoptosis induction. Additionally, cytotoxicity assays were performed to evaluate cell viability and proliferation, confirming the dose-dependent effects of the treatments on apoptosis and cancer cell inhibition.

Results

The IC₅₀ values for the treatment were 389.7 µg/mL and 297.1 µg/mL at 24 and 48 hr, respectively, significantly increasing the pro-apoptotic gene CASP9 while decreasing CYCLIND1, MMP2, and VEGF expression. GCNPs were synthesized and characterized, showing cytotoxic effects with IC₅₀ values of 21 µg/mL at 24 hr and 9.859 µg/mL at 48 hr. These nanoparticles enhanced apoptosis by increasing Bax expression and decreasing Bcl-2, while also suppressing MMP2 and MMP9, which are linked to metastasis. Chitosan nanoparticles containing EGCG demonstrated notable anti-cancer effects, reducing cell proliferation and inducing apoptosis. Additionally, Lactobacillus strains from Chlorella vulgaris exhibited strong probiotic and anti-cancer properties. Cytotoxicity assays revealed a dose-dependent inhibition of Panc-1 cell viability and induction of apoptosis, highlighting the potential of these natural compounds in cancer therapy.

Conclusion

In conclusion, the study reveals that Lactobacillus buchneri supernatant, chitosan nanoparticles conjugated with genistein, and chitosan nanoparticles containing EGCG significantly induce apoptosis in Panc-1 pancreatic cancer cells. The IC₅₀ values indicate strong cytotoxic effects, particularly with GCNPs. The upregulation of CASP9 and downregulation of Bcl-2 highlight the treatments' potential to promote apoptosis. These findings emphasize the promising role of natural compounds and nanotechnology in developing effective pancreatic cancer therapies.