Lacticaseibacillus casei and Lacticaseibacillus rhamnosus cell-free supernatants induce apoptosis-related responses in PANC-1 human pancreatic cancer cells
-
Shadi Moradi
,
Reza Moradi-Hajidavaloo
,
Fatemeh Soleimanifar
,
Kiana Shahzamani
,
Mehdi Hajian
,
Hajar-Alsadat Mansouri-Tehrani
Abstract
Background and Objectives: This study aimed to evaluate the anticancer potential of Lacticaseibacillus casei and Lacticaseibacillus rhamnosus cell-free supernatants (CFSs) against the PANC-1 human pancreatic cancer cell line, focusing on apoptosis, cell cycle modulation, and the expression of BAX and BCL-XL genes.
Materials and Methods: PANC-1 cancer cells and adult human dermal fibroblast (HDFa) cells were treated with various concentrations of individual or combined CFSs. Cell viability was assessed using the MTT assay. Apoptosis was evaluated through Hoechst/PI staining and flow cytometry, while cell cycle distribution was analyzed via flow cytometry. Gene expression of BAX and BCL-XL was measured by quantitative real-time PCR.
Results: At 20% (v/v), all CFS treatments significantly reduced PANC-1 cell viability while showing minimal effects on HDFa cells. Flow cytometry confirmed apoptotic rates of 39.33%, 42%, and 40.33% for L. casei, L. rhamnosus, and their combination, respectively, alongside a notable increase in S-phase cell population. Gene expression analysis showed a pro-apoptotic shift, characterized by BAX upregulation and BCL-XL downregulation.
Conclusion: CFSs from L. casei and L. rhamnosus showed anticancer effects on PANC-1 cells, inducing apoptosis, S-phase arrest, and a favorable shift in apoptosis-related gene expression. These findings highlight their potential as promising adjuvant candidates for pancreatic cancer therapy.
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11. Thilakarathna WP, Rupasinghe HV, Ridgway ND. Mechanisms by which probiotic bacteria attenuate the risk of hepatocellular carcinoma. Int J Mol Sci 2021; 22: 2606.
12. Ravi AV, Musthafa KS, Jegathammbal G, Kathiresan K, Pandian SK. Screening and evaluation of probiotics as a biocontrol agent against pathogenic Vibrios in marine aquaculture. Lett Appl Microbiol 2007; 45: 219-223.
13. Uccello M, Malaguarnera G, Basile F, D’agata V, Malaguarnera M, Bertino G, et al. Potential role of probiotics on colorectal cancer prevention. BMC Surg 2012; 12(Suppl 1): S35.
14. Lamichhane P, Maiolini M, Alnafoosi O, Hussein S, Alnafoosi H, Umbela S, et al. Colorectal cancer and probiotics: are bugs really drugs? Cancers (Basel) 2020; 12: 1162.
15. Motevaseli E, Dianatpour A, Ghafouri-Fard S. The role of probiotics in cancer treatment: emphasis on their in vivo and in vitro anti-metastatic effects. Int J Mol Cell Med 2017; 6: 66-76.
16. Shenderov BA. Metabiotics: novel idea or natural development of probiotic conception. Microb Ecol Health Dis 2013; 24. doi: 10.3402/mehd.v24i0.20399. eCollection 2013
17. Nataraj BH, Ali SA, Behare PV, Yadav H. Postbiotics-parabiotics: The new horizons in microbial biotherapy and functional foods. Microb Cell Fact 2020; 19: 168.
18. Konishi H, Fujiya M, Tanaka H, Ueno N, Moriichi K, Sasajima J, et al. Probiotic-derived ferrichrome inhibits colon cancer progression via JNK-mediated apoptosis. Nat Commun 2016; 7: 12365.
19. Kwoji ID, Aiyegoro OA, Okpeku M, Adeleke MA. Multi-strain probiotics: synergy among isolates enhances biological activities. Biology (Basel) 2021; 10: 322.
20. Miyaguchi J, Shiga K, Ogawa K, Suzuki F, Katagiri K, Saito D, et al. Treatment with Lactobacillus retards the tumor growth of head and neck squamous cell carcinoma cells inoculated in mice. Tohoku J Exp Med 2018; 245: 269-275.
21. Tiptiri-Kourpeti A, Spyridopoulou K, Santarmaki V, Aindelis G, Tompoulidou E, Lamprianidou EE, et al. Lactobacillus casei exerts anti-proliferative effects accompanied by apoptotic cell death and up-regulation of TRAIL in colon carcinoma cells. PloS One 2016; 11(2): e0147960.
22. Dolati M, Tafvizi F, Salehipour M, Movahed TK, Jafari P. Inhibitory effects of probiotic Bacillus coagulans against MCF7 breast cancer cells. Iran J Microbiol 2021; 13: 839-847.
23. Bayoumi MA, Griffiths MW. In vitro inhibition of expression of virulence genes responsible for colonization and systemic spread of enteric pathogens using Bifidobacterium bifidum secreted molecules. Int J Food Microbiol 2012; 156: 255-263.
24. Orlando A, Messa C, Linsalata M, Cavallini A, Russo F. Effects of Lactobacillus rhamnosus GG on proliferation and polyamine metabolism in HGC-27 human gastric and DLD-1 colonic cancer cell lines. Immunopharmacol Immunotoxicol 2009; 31: 108-116.
25. Sadeghi-aliabadi H, Mohammadi F, Fazeli H, Mirlohi M. Effects of Lactobacillus plantarum A7 with probiotic potential on colon cancer cell proliferation in comparison with a commercial strain. Iran J Basic Med Sci 2014; 17: 815-819.
26. Choi SS, Kim Y, Han KS, You S, Oh S, Kim SH. Effects of Lactobacillus strains on cancer cell proliferation and oxidative stress in vitro. Lett Appl Microbiol 2006; 42: 452-458.
27. Lau AC, Cui H, Tymianski M. The use of propidium iodide to assess excitotoxic neuronal death in primary mixed cortical cultures. Methods Mol Biol 2007; 399: 15-29.
28. Sankarapandian V, Venmathi Maran BA, Rajendran RL, Jogalekar MP, Gurunagarajan S, Krishnamoorthy R, et al. An update on the effectiveness of probiotics in the prevention and treatment of cancer. Life (Basel) 2022; 12: 59.
29. Erdem SA, Nabavi SF, Orhan IE, Daglia M, Izadi M, Nabavi SM. Blessings in disguise: a review of phytochemical composition and antimicrobial activity of plants belonging to the genus Eryngium. Daru 2015; 23: 53.
30. Nami Y, Abdullah N, Haghshenas B, Radiah D, Rosli R, Khosroushahi AY. Assessment of probiotic potential and anticancer activity of newly isolated vaginal bacterium Lactobacillus plantarum 5BL. Microbiol Immunol 2014; 58: 492-502.
31. Malik SS, Saeed A, Baig M, Asif N, Masood N, Yasmin A. Anticarcinogenecity of microbiota and probiotics in breast cancer. Int J Food Prop 2018; 21: 655-666.
32. Rajoka MS, Zhao H, Lu Y, Lian Z, Li N, Hussain N, et al. Anticancer potential against cervix cancer (HeLa) cell line of probiotic Lactobacillus casei and Lactobacillus paracasei strains isolated from human breast milk. Food Funct 2018; 9: 2705-2715.
33. Kita A, Fujiya M, Konishi H, Tanaka H, Kashima S, Iwama T, et al. Probiotic-derived ferrichrome inhibits the growth of refractory pancreatic cancer cells. Int J Oncol 2020; 57: 721-732.
34. Gantois I, Ducatelle R, Pasmans F, Haesebrouck F, Hautefort I, Thompson A, et al. Butyrate specifically down-regulates Salmonella pathogenicity island 1 gene expression. Appl Environ Microbiol 2006; 72: 946-949.
35. Kim JY, Woo HJ, Kim YS, Kim KH, Lee HJ. Cell cycle dysregulation induced by cytoplasm of Lactococcus lactis ssp. lactis in SNUC2A, a colon cancer cell line. Nutr Cancer 2003; 46: 197-201.
36. Ma EL, Choi YJ, Choi J, Pothoulakis C, Rhee SH, Im E. The anticancer effect of probiotic Bacillus polyfermenticus on human colon cancer cells is mediated through ErbB2 and ErbB3 inhibition. Int J Cancer 2010; 127: 780-790.
37. Sharma M, Shukla G. Metabiotics: one step ahead of probiotics; an insight into mechanisms involved in anticancerous effect in colorectal cancer. Front Microbiol 2016; 7: 1940.
38. Azam R, Ghafouri-Fard S, Tabrizi M, Modarressi MH, Ebrahimzadeh-Vesal R, Daneshvar M, et al. Lactobacillus acidophilus and Lactobacillus crispatus culture supernatants downregulate expression of cancer-testis genes in the MDA-MB-231 cell line. Asian Pac J Cancer Prev 2014; 15: 4255-4259.
39. Chumchalová J, Šmarda J. Human tumor cells are selectively inhibited by colicins. Folia Microbiol (Basel) 2003; 48: 111-115.
40. Zhang YJ, Li S, Gan RY, Zhou T, Xu DP, Li HB. Impacts of gut bacteria on human health and diseases. Int J Mol Sci 2015; 16: 7493-7519.
41. Feng Q, Chen WD, Wang YD. Gut microbiota: an integral moderator in health and disease. Front Microbiol 2018; 9: 151.
42. Dehghani N, Tafvizi F, Jafari P. Cell cycle arrest and anti-cancer potential of probiotic Lactobacillus rhamnosus against HT-29 cancer cells. BioImpacts 2020; 11: 245-252.
43. Diep DB, Mathiesen G, Eijsink VG, Nes IF. Use of lactobacilli and their pheromone-based regulatory mechanism in gene expression and drug delivery. Curr Pharm Biotechnol 2009; 10: 62-73.
44. Maldonado A, Ruiz-Barba JL, Jiménez-Díaz R. Production of plantaricin NC8 by Lactobacillus plantarum NC8 is induced in the presence of different types of gram-positive bacteria. Arch Microbiol 2004; 181: 8-16.
45. Louis P, Hold GL, Flint HJ. The gut microbiota, bacterial metabolites and colorectal cancer. Nat Rev Microbiol 2014; 12: 661-672.
2. Tsilingiri K, Barbosa T, Penna G, Caprioli F, Sonzogni A, Viale G, et al. Probiotic and postbiotic activity in health and disease: comparison on a novel polarised ex-vivo organ culture model. Gut 2012; 61: 1007-1015.
3. Panebianco C, Latiano T, Pazienza V. Microbiota manipulation by probiotics administration as emerging tool in cancer prevention and therapy. Front Oncol 2020; 10: 679.
4. Aagaard K, Ma J, Antony KM, Ganu R, Petrosino J, Versalovic J. The placenta harbors a unique microbiome. Sci Transl Med 2014; 6(237): 237ra65.
5. Plaza-Diaz J, Ruiz-Ojeda FJ, Gil-Campos M, Gil A. Mechanisms of action of probiotics. Adv Nutr 2019; 10(suppl_1), S49-S66.
6. Ammer-Herrmenau C, Pfisterer N, Weingarten MF, Neesse A. The microbiome in pancreatic diseases: Recent advances and future perspectives. United European Gastroenterol J 2020; 8: 878-885.
7. Śliżewska K, Markowiak-Kopeć P, Śliżewska W. The role of probiotics in cancer prevention. Cancers (Basel) 2020; 13: 20.
8. Leal-Lopes C, Velloso FJ, Campopiano JC, Sogayar MC, Correa RG. Roles of commensal microbiota in pancreas homeostasis and pancreatic pathologies. J Diabetes Res 2015; 2015: 284680.
9. Kim BK, Yoon YS, Ryu Y, Chung MJ. Probiotic-derived p8 protein induce apoptosis via regulation of RNF152 in colorectal cancer cells. Am J Cancer Res 2021; 11: 746-759.
10. Prazdnova EV, Mazanko MS, Chistyakov VA, Bogdanova AA, Refeld AG, Kharchenko EY, et al. Antimutagenic Activity as a Criterion of Potential Probiotic Properties. Probiotics Antimicrob Proteins 2022; 14: 1094-1109.
11. Thilakarathna WP, Rupasinghe HV, Ridgway ND. Mechanisms by which probiotic bacteria attenuate the risk of hepatocellular carcinoma. Int J Mol Sci 2021; 22: 2606.
12. Ravi AV, Musthafa KS, Jegathammbal G, Kathiresan K, Pandian SK. Screening and evaluation of probiotics as a biocontrol agent against pathogenic Vibrios in marine aquaculture. Lett Appl Microbiol 2007; 45: 219-223.
13. Uccello M, Malaguarnera G, Basile F, D’agata V, Malaguarnera M, Bertino G, et al. Potential role of probiotics on colorectal cancer prevention. BMC Surg 2012; 12(Suppl 1): S35.
14. Lamichhane P, Maiolini M, Alnafoosi O, Hussein S, Alnafoosi H, Umbela S, et al. Colorectal cancer and probiotics: are bugs really drugs? Cancers (Basel) 2020; 12: 1162.
15. Motevaseli E, Dianatpour A, Ghafouri-Fard S. The role of probiotics in cancer treatment: emphasis on their in vivo and in vitro anti-metastatic effects. Int J Mol Cell Med 2017; 6: 66-76.
16. Shenderov BA. Metabiotics: novel idea or natural development of probiotic conception. Microb Ecol Health Dis 2013; 24. doi: 10.3402/mehd.v24i0.20399. eCollection 2013
17. Nataraj BH, Ali SA, Behare PV, Yadav H. Postbiotics-parabiotics: The new horizons in microbial biotherapy and functional foods. Microb Cell Fact 2020; 19: 168.
18. Konishi H, Fujiya M, Tanaka H, Ueno N, Moriichi K, Sasajima J, et al. Probiotic-derived ferrichrome inhibits colon cancer progression via JNK-mediated apoptosis. Nat Commun 2016; 7: 12365.
19. Kwoji ID, Aiyegoro OA, Okpeku M, Adeleke MA. Multi-strain probiotics: synergy among isolates enhances biological activities. Biology (Basel) 2021; 10: 322.
20. Miyaguchi J, Shiga K, Ogawa K, Suzuki F, Katagiri K, Saito D, et al. Treatment with Lactobacillus retards the tumor growth of head and neck squamous cell carcinoma cells inoculated in mice. Tohoku J Exp Med 2018; 245: 269-275.
21. Tiptiri-Kourpeti A, Spyridopoulou K, Santarmaki V, Aindelis G, Tompoulidou E, Lamprianidou EE, et al. Lactobacillus casei exerts anti-proliferative effects accompanied by apoptotic cell death and up-regulation of TRAIL in colon carcinoma cells. PloS One 2016; 11(2): e0147960.
22. Dolati M, Tafvizi F, Salehipour M, Movahed TK, Jafari P. Inhibitory effects of probiotic Bacillus coagulans against MCF7 breast cancer cells. Iran J Microbiol 2021; 13: 839-847.
23. Bayoumi MA, Griffiths MW. In vitro inhibition of expression of virulence genes responsible for colonization and systemic spread of enteric pathogens using Bifidobacterium bifidum secreted molecules. Int J Food Microbiol 2012; 156: 255-263.
24. Orlando A, Messa C, Linsalata M, Cavallini A, Russo F. Effects of Lactobacillus rhamnosus GG on proliferation and polyamine metabolism in HGC-27 human gastric and DLD-1 colonic cancer cell lines. Immunopharmacol Immunotoxicol 2009; 31: 108-116.
25. Sadeghi-aliabadi H, Mohammadi F, Fazeli H, Mirlohi M. Effects of Lactobacillus plantarum A7 with probiotic potential on colon cancer cell proliferation in comparison with a commercial strain. Iran J Basic Med Sci 2014; 17: 815-819.
26. Choi SS, Kim Y, Han KS, You S, Oh S, Kim SH. Effects of Lactobacillus strains on cancer cell proliferation and oxidative stress in vitro. Lett Appl Microbiol 2006; 42: 452-458.
27. Lau AC, Cui H, Tymianski M. The use of propidium iodide to assess excitotoxic neuronal death in primary mixed cortical cultures. Methods Mol Biol 2007; 399: 15-29.
28. Sankarapandian V, Venmathi Maran BA, Rajendran RL, Jogalekar MP, Gurunagarajan S, Krishnamoorthy R, et al. An update on the effectiveness of probiotics in the prevention and treatment of cancer. Life (Basel) 2022; 12: 59.
29. Erdem SA, Nabavi SF, Orhan IE, Daglia M, Izadi M, Nabavi SM. Blessings in disguise: a review of phytochemical composition and antimicrobial activity of plants belonging to the genus Eryngium. Daru 2015; 23: 53.
30. Nami Y, Abdullah N, Haghshenas B, Radiah D, Rosli R, Khosroushahi AY. Assessment of probiotic potential and anticancer activity of newly isolated vaginal bacterium Lactobacillus plantarum 5BL. Microbiol Immunol 2014; 58: 492-502.
31. Malik SS, Saeed A, Baig M, Asif N, Masood N, Yasmin A. Anticarcinogenecity of microbiota and probiotics in breast cancer. Int J Food Prop 2018; 21: 655-666.
32. Rajoka MS, Zhao H, Lu Y, Lian Z, Li N, Hussain N, et al. Anticancer potential against cervix cancer (HeLa) cell line of probiotic Lactobacillus casei and Lactobacillus paracasei strains isolated from human breast milk. Food Funct 2018; 9: 2705-2715.
33. Kita A, Fujiya M, Konishi H, Tanaka H, Kashima S, Iwama T, et al. Probiotic-derived ferrichrome inhibits the growth of refractory pancreatic cancer cells. Int J Oncol 2020; 57: 721-732.
34. Gantois I, Ducatelle R, Pasmans F, Haesebrouck F, Hautefort I, Thompson A, et al. Butyrate specifically down-regulates Salmonella pathogenicity island 1 gene expression. Appl Environ Microbiol 2006; 72: 946-949.
35. Kim JY, Woo HJ, Kim YS, Kim KH, Lee HJ. Cell cycle dysregulation induced by cytoplasm of Lactococcus lactis ssp. lactis in SNUC2A, a colon cancer cell line. Nutr Cancer 2003; 46: 197-201.
36. Ma EL, Choi YJ, Choi J, Pothoulakis C, Rhee SH, Im E. The anticancer effect of probiotic Bacillus polyfermenticus on human colon cancer cells is mediated through ErbB2 and ErbB3 inhibition. Int J Cancer 2010; 127: 780-790.
37. Sharma M, Shukla G. Metabiotics: one step ahead of probiotics; an insight into mechanisms involved in anticancerous effect in colorectal cancer. Front Microbiol 2016; 7: 1940.
38. Azam R, Ghafouri-Fard S, Tabrizi M, Modarressi MH, Ebrahimzadeh-Vesal R, Daneshvar M, et al. Lactobacillus acidophilus and Lactobacillus crispatus culture supernatants downregulate expression of cancer-testis genes in the MDA-MB-231 cell line. Asian Pac J Cancer Prev 2014; 15: 4255-4259.
39. Chumchalová J, Šmarda J. Human tumor cells are selectively inhibited by colicins. Folia Microbiol (Basel) 2003; 48: 111-115.
40. Zhang YJ, Li S, Gan RY, Zhou T, Xu DP, Li HB. Impacts of gut bacteria on human health and diseases. Int J Mol Sci 2015; 16: 7493-7519.
41. Feng Q, Chen WD, Wang YD. Gut microbiota: an integral moderator in health and disease. Front Microbiol 2018; 9: 151.
42. Dehghani N, Tafvizi F, Jafari P. Cell cycle arrest and anti-cancer potential of probiotic Lactobacillus rhamnosus against HT-29 cancer cells. BioImpacts 2020; 11: 245-252.
43. Diep DB, Mathiesen G, Eijsink VG, Nes IF. Use of lactobacilli and their pheromone-based regulatory mechanism in gene expression and drug delivery. Curr Pharm Biotechnol 2009; 10: 62-73.
44. Maldonado A, Ruiz-Barba JL, Jiménez-Díaz R. Production of plantaricin NC8 by Lactobacillus plantarum NC8 is induced in the presence of different types of gram-positive bacteria. Arch Microbiol 2004; 181: 8-16.
45. Louis P, Hold GL, Flint HJ. The gut microbiota, bacterial metabolites and colorectal cancer. Nat Rev Microbiol 2014; 12: 661-672.
| Files | ||
| Issue | Vol 18 No 3 (2026) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v18i3.21651 | |
| Keywords | ||
| Lacticaseibacillus casei Lacticaseibacillus rhamnosus Pancreatic cancer BAX BCL-XL | ||
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How to Cite
1.
Moradi S, Moradi-Hajidavaloo R, Soleimanifar F, Shahzamani K, Hajian M, Mansouri-Tehrani H-A. Lacticaseibacillus casei and Lacticaseibacillus rhamnosus cell-free supernatants induce apoptosis-related responses in PANC-1 human pancreatic cancer cells. Iran J Microbiol. 2026;18(3):295-305.
