Isolated Lactobacillus fermentum Ab.RS22 from traditional dairy products inhibits HeLa cervical cancer cell proliferation and modulates apoptosis by the PTEN-Akt pathway

Document Type : Original Article

Authors

1 INSERM U1148, Laboratory for Vascular Translation Science (LVTS), Cardiovascular Bioengineering, University Sorbonne Paris North, Paris, France

2 Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran

3 Clinical Research Development Unit, Booalisina Hospital, Qazvin University of Medical Sciences, Qazvin, Iran

4 Student Research Committee, Qazvin University of Medical Sciences, Qazvin, Iran

5 Clinical Pharmacist, Baylor Scott & White Medical Center – Lakeway 100 Medical Pkwy, Lakeway, TX 78738

6 Rayan Novin Pajoohan Pras, Biotechnology Company, Biotechnology Incubator, Shiraz University of Medical Sciences, Shiraz, Iran

7 Medical Microbiology Research Center, Qazvin University of Medical Sciences, Qazvin, Iran

8 Department of Medical Parasitology and Mycology, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran

9 Department of Biochemistry and Genetics, School of Medicine, Qazvin University of Medical Sciences, Qazvin, Iran

10.22038/ijbms.2023.72825.15846

Abstract

Objective(s): It is worthwhile to note that, some probiotics such as Lactobacilli and Bifidobacteria isolated from dairy products have significant therapeutic effects against cancer cells. Here, we evaluated anti-proliferation and the apoptotic effects of isolated Lactobacillus fermentum Ab.RS22 from traditional dairy products on the HeLa cervical cancer cells in vitro.
Materials and Methods: The viability of treated HeLa cells with supernatant of Lactobacillus in 0.5, 0.75, 1, 1.5, and 2 ng/ml concentrations, and IC50 values were detected by tetrazolium bromide. The L. fermentum Ab.RS22-induced cell death by flow cytometry was confirmed through evaluation of the expression of caspase-3, P53, PTEN, and AKT genes by quantitative reverse transcription-polymerase chain reactions (qRT-PCR).
Results: Most cytotoxicity effects of Lactobacillus on HeLa cells were detected in 2 ng/ml at 24 hr (P<0.01); also, the IC50 value was measured as 1.5 ng/ml. The findings of the flow cytometry assay showed that L. fermentum Ab.RS22 in 1.5 ng/ml concentration at 24 hr increased the percentage of both apoptosis and necrosis cells. Lactobacillus-induced cell death was verified through results of Real-time PCR; where expression of caspase-3, P53, and PTEN genes was increased (P<0.01), and also expression of AKT gene (anti-apoptotic) was decreased (P<0.05). 
Conclusion: Our findings showed that L. fermentum Ab.RS22 could dose-dependently inhibit the proliferation of the HeLa cells. Its apoptotic effect was confirmed via modulating PTEN/p53/Akt gene expression and activation of the caspase-3 mediated apoptosis pathway. Therefore, L. fermentum Ab.RS22 can be considered a valuable anticancer candidate against cervical cancer progression in subsequent studies. 

Keywords

Main Subjects


1. Bañuelos-Villegas EG, Pérez-yPérez MF, Alvarez-Salas LM. Cervical cancer, papillomavirus, and miRNA dysfunction. Front Mol Biosci 2021; 8: 758-771. 
2. Farahmand M, Moghoofei M, Dorost A, Abbasi S, Monavari SH, Kiani SJ, Tavakoli A. Prevalence and genotype distribution of genital human papillomavirus infection in female sex workers in the world: A systematic review and meta-analysis. BMC Public Health 2020; 20: 1455-1469. 
3. Ferrall L, Lin KY, Roden RBS, Hung CF, Wu TC.Cervical cancer immunotherapy: facts and hopes. Clin Cancer Res 2021; 27: 4953-4973. 
4. Zhou ZW, Long HZ, Cheng Y, Luo HY, Wen DD, Gao LC .From microbiome to inflammation: The key drivers of cervical cancer. Front Microbiol 2021; 12: 767-931. 
5. Jahanshahi M, Maleki Dana P, Badehnoosh B, Asemi Z, Hallajzadeh J, Mansournia MA, et al. Anti-tumor activities of probiotics in cervical cancer. J Ovarian Res 2020; 13: 68-79. 
6. Frąszczak K, Barczyński B, Kondracka A. Does Lactobacillus exert a protective effect on the development of cervical and endometrial cancer in women. Cancers  2022; 14: 4909-4936. 
7. Yang X, Da M, Zhang W, Qi Q, Zhang C, Han S. Role of  Lactobacillus in cervical cancer. Cancer Manag Res 2018; 10:1219-1229. 
8. Ikeda Y, Adachi K, Tomio K, Eguchi-Kojima S, Tsuruga T, Uchino-Mori M, et al.A placebo-controlled, double-blind randomized (phase IIB) trial of oral administration with HPV16 E7-expressing Lactobacillus, GLBL101c, for the treatment of cervical intraepithelial neoplasia grade 2 (CIN2). Vaccines 2021; 9: 329-342. 
9. Gamallat Y, Meyiah A, Kuugbee ED, Hago AM, Chiwala G, Awadasseid A, et al. Lactobacillus rhamnosus induced epithelial cell apoptosis, ameliorates inflammation and prevents colon cancer development in an animal model. Biomed Pharmacother 2016; 83: 536-541. 
10. Maleki-Kakelar H, Dehghani J, Barzegari A, Barar J, Shirmohamadi M, Sadeghi J, et al. Lactobacillus plantarum induces apoptosis in gastric cancer cells via modulation of signaling pathways in Helicobacter pylori. BioImpacts  2020;10: 65-72. 
11. Li N, Men W, Zheng Y, Wang H, Meng X. Oroxin B induces apoptosis by down-regulating microRNA-221 resulting in the inactivation of the PTEN/PI3K/AKT pathway in liver cancer. Molecules 2019; 24: 4384-4399. 
12. Ztaliou I, Tsakalidou E, Tzanetakis N, Kalantzopoulos G. Lactobacillus plantarum strains isolated from traditional Greek cheese. Taxonomic characterization and screening for enzyme activities. Lait  1996; 76: 209-216. 
13. Asoudeh-Fard A, Barzegari A, Dehnad A, Bastani S, Golchin A, Omidi Y. Lactobacillus plantarum induces apoptosis in oral cancer KB cells through upregulation of PTEN and downregulation of MAPK signalling pathways. BioImpacts 2017; 7: 193-198. 
14. Zhang T, Pan D, Yang Y, Jiang X, Zhang J, Zeng X, et al . Effect of Lactobacillus acidophilus CICC 6074 S-Layer Protein on colon cancer HT-29 cell proliferation and apoptosis. J Agric Food Chem 2020; 68: 2639-2647. 
15. Salehi M, Piri H, Farasat A, Pakbin B, Gheibi N. Activation of apoptosis and G0/G1 cell cycle arrest along with inhibition of melanogenesis by humic acid and fulvic acid: BAX/BCL-2 and Tyr genes expression and evaluation of nanomechanical properties in A375 human melanoma cell line. Iran J Basic Med Sci 2022; 25: 489-496. 
16. De Loera Rodríguez LH, Ortiz GG, Rivero Moragrega P, Velázquez Brizuela IE, Santoscoy Gutiérrez JF, Rincón Sánchez AR, et al. Effect of symbiotic supplementation on fecal calprotectin levels and lactic acid bacteria, Bifidobacteria, Escherichia coli and Salmonella DNA in patients with cervical cancer. Nutr Hosp 2018; 35: 1394-1400. 
17. Li Y, Yu T, Yan H, Li D, Yu T, Yuan T, et al. Vaginal microbiota and HPV infection: Novel mechanistic insights and therapeutic strategies. Infect Drug Resist 2020;13: 1213-1220. 
18. Qiu G, Yu Y, Wang Y, Wang X. The significance of probiotics in preventing radiotherapy-induced diarrhea in patients with cervical cancer: A systematic review and meta-analysis. Int J Surg 2019; 65: 61-69. 
19. Abdolalipour E, Mahooti M, Gorji A, Ghaemi A. Synergistic therapeutic effects of probiotic Lactobacillus casei TD-2 consumption on GM-CSF-Induced Immune Responses in a Murine Model of Cervical Cancer. Nutr Cancer  2022; 74: 372-382. 
20. Abdolalipour E, Mahooti M, Salehzadeh A, Torabi A, Mohebbi SR, Gorji A, et al. Evaluation of the antitumor immune responses of probiotic Bifidobacterium bifidum in human papillomavirus-induced tumor model. Microb Pathog  2020; 145: 104-116. 
21. Isazadeh A, Hajazimian S, Shadman B, Safaei S, Babazadeh Bedoustani A, Chavoshi R, et al. Anti-cancer effects of probiotic Lactobacillus acidophilus for colorectal cancer cell Line Caco-2 through apoptosis induction. Pharm Sci 2021; 27:262-267. 
22. Sadek KM, Lebda MA, Nasr NE, Nasr SM, El-Sayed Y. Role of lncRNAs as prognostic markers of hepatic cancer and potential therapeutic targeting by S-adenosylmethionine via inhibiting PI3K/Akt signaling pathways. Environ Sci Pollut Res Int 2018;25:20057-20070. 
23. Moradzadeh M, Hosseini A, Erfanian S, Rezaei H. Epigallocatechin-3-gallate promotes apoptosis in human breast cancer T47D cells through down-regulation of PI3K/AKT and telomerase. Pharmacol Rep 2017; 69 : 924-932. 
24. Sadek KM, Abouzed TK, Abouelkhair R, Nasr S. The chemo-prophylactic efficacy of an ethanol Moringa oleifera leaf extract against hepatocellular carcinoma in rats. Pharm Biol 2017; 55 :1458-1466. 
25. Rahbar Saadat Y, Pourghassem Gargari B, Shahabi A, Nami Y, Yari Khosroushahi A. Prophylactic role of Lactobacillus paracasei exopolysaccharides on colon cancer cells through apoptosis not ferroptosis. Pharm Sci 2021; 27: 251-261. 
26. Zununi Vahed S, Barzegari A, Rahbar Saadat Y, Goreyshi A, Omidi Y. Leuconostoc mesenteroides-derived anticancer pharmaceuticals hinder inflammation and cell survival in colon cancer cells by modulating NF-κB/AKT/PTEN/MAPK pathways. Biomed Pharmacother 2017; 94:1094-1100. 
27. SungurT, Aslim B, Karaaslan C, Aktas B. Impact of Exopolysaccharides (EPSs) of Lactobacillus gasseri strains isolated from human vagina on cervical tumor cells (HeLa). Anaerobe 2017; 47: 137-144.