Assessment of Cell Cycle and Induction of Apoptosis by Shiga-like Toxin Produced by Escherichia coli O157:H7 in T47D Breast Cancer Cells Using Flow Cytometry
Background: The low general toxicity against tumors expressing globotriaosylceramide (Gb3) and Shiga-like toxins produced by E. coli have been proposed as an anti-cancer therapy because of their specific target. This study aimed to determine the potency of the local strains of E. coli O157:H7 isola...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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PubMed Central PMC
2022
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| Online Access: | https://repository.ugm.ac.id/281800/1/Widiasih_KH.pdf |
| _version_ | 1826050435972071424 |
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| author | Suardana, I Wayan Swacita, Ida Bagus Ngurah Pinatih, Komang Januartha Putra Suharsono, Hamong Widiasih, Dyah Ayu |
| author_facet | Suardana, I Wayan Swacita, Ida Bagus Ngurah Pinatih, Komang Januartha Putra Suharsono, Hamong Widiasih, Dyah Ayu |
| author_sort | Suardana, I Wayan |
| collection | UGM |
| description | Background: The low general toxicity against tumors expressing globotriaosylceramide (Gb3) and Shiga-like toxins
produced by E. coli have been proposed as an anti-cancer therapy because of their specific target. This study aimed to determine the potency of the local strains of E. coli O157:H7 isolated from humans and cattle as a new breast cancer therapy by analyzing the cell cycle’s inhibition and apoptosis induction. Material and Methods: Approximately 10
cultured T47D cells were subjected to Shiga-like toxin produced by four local isolates of E. coli O157:H7, including KL-48 (2) from humans, and SM-25 (1), SM-7 (1), DS-21 (4) from cattle. Using ATCC 43894 as a control, the treatment was observed for 24 h by two replications. In addition, a FITC-Annexin V and PI assay were used to observe apoptosis and necrosis effect, as well as to analyze the cell cycle using propidium iodide (PI) staining. Results: The results showed the toxicity effect of Shiga in the human T47 D cells line. The viability of the cells is subjected to Shiga-like toxins produced by KL-48 (2), SM7 (1), ATCC 43894, SM-25 (1), and DS-21 (4) isolates decreased with 15.20, 16.36, 22.17, 22.64, and 33.86%, in contrary to control of 94.36%. These were supported by the cells entering the late apoptosis of the cell cycle through each isolate with 67.66, 62.60, 63.68, 63.90, and 54.74%, and a control of 0.01%. Also, the necrosis cell for each treatment of 12.73, 19.3, 10.84, 10.53, and 4.86% was higher than the control of 5.51%. These were confirmed by the higher percentage of the cells treated with toxins of KL-48 (2), SM7(1), ATCC 43894, SM-25 (1), and DS-21 (4), which entered G0-G1 of the cell cycle phase with 66.41, 63.37, 61.52, 55.36, and 47.28%, respectively, than control of 40.69%. Additionally, the toxicity effect was supported by an increase in the cells entering the S and the G2-M phase of the cycle for each treatment. Conclusion: It is concluded that the Shiga-like toxin produced by E. coli O157:H7 local isolates can be developed as a drug against breast cancer based on its effect to arrest induction of the cell cycle and inducing apoptosis. |
| first_indexed | 2024-03-14T00:03:56Z |
| format | Article |
| id | oai:generic.eprints.org:281800 |
| institution | Universiti Gadjah Mada |
| language | English |
| last_indexed | 2024-03-14T00:03:56Z |
| publishDate | 2022 |
| publisher | PubMed Central PMC |
| record_format | dspace |
| spelling | oai:generic.eprints.org:2818002023-11-13T07:05:25Z https://repository.ugm.ac.id/281800/ Assessment of Cell Cycle and Induction of Apoptosis by Shiga-like Toxin Produced by Escherichia coli O157:H7 in T47D Breast Cancer Cells Using Flow Cytometry Suardana, I Wayan Swacita, Ida Bagus Ngurah Pinatih, Komang Januartha Putra Suharsono, Hamong Widiasih, Dyah Ayu Veterinary Sciences Background: The low general toxicity against tumors expressing globotriaosylceramide (Gb3) and Shiga-like toxins produced by E. coli have been proposed as an anti-cancer therapy because of their specific target. This study aimed to determine the potency of the local strains of E. coli O157:H7 isolated from humans and cattle as a new breast cancer therapy by analyzing the cell cycle’s inhibition and apoptosis induction. Material and Methods: Approximately 10 cultured T47D cells were subjected to Shiga-like toxin produced by four local isolates of E. coli O157:H7, including KL-48 (2) from humans, and SM-25 (1), SM-7 (1), DS-21 (4) from cattle. Using ATCC 43894 as a control, the treatment was observed for 24 h by two replications. In addition, a FITC-Annexin V and PI assay were used to observe apoptosis and necrosis effect, as well as to analyze the cell cycle using propidium iodide (PI) staining. Results: The results showed the toxicity effect of Shiga in the human T47 D cells line. The viability of the cells is subjected to Shiga-like toxins produced by KL-48 (2), SM7 (1), ATCC 43894, SM-25 (1), and DS-21 (4) isolates decreased with 15.20, 16.36, 22.17, 22.64, and 33.86%, in contrary to control of 94.36%. These were supported by the cells entering the late apoptosis of the cell cycle through each isolate with 67.66, 62.60, 63.68, 63.90, and 54.74%, and a control of 0.01%. Also, the necrosis cell for each treatment of 12.73, 19.3, 10.84, 10.53, and 4.86% was higher than the control of 5.51%. These were confirmed by the higher percentage of the cells treated with toxins of KL-48 (2), SM7(1), ATCC 43894, SM-25 (1), and DS-21 (4), which entered G0-G1 of the cell cycle phase with 66.41, 63.37, 61.52, 55.36, and 47.28%, respectively, than control of 40.69%. Additionally, the toxicity effect was supported by an increase in the cells entering the S and the G2-M phase of the cycle for each treatment. Conclusion: It is concluded that the Shiga-like toxin produced by E. coli O157:H7 local isolates can be developed as a drug against breast cancer based on its effect to arrest induction of the cell cycle and inducing apoptosis. PubMed Central PMC 2022 Article PeerReviewed application/pdf en https://repository.ugm.ac.id/281800/1/Widiasih_KH.pdf Suardana, I Wayan and Swacita, Ida Bagus Ngurah and Pinatih, Komang Januartha Putra and Suharsono, Hamong and Widiasih, Dyah Ayu (2022) Assessment of Cell Cycle and Induction of Apoptosis by Shiga-like Toxin Produced by Escherichia coli O157:H7 in T47D Breast Cancer Cells Using Flow Cytometry. Asian Pacific Journal of Cancer Prevention, 23 (10). pp. 3247-3252. ISSN 2476-762X https://journal.waocp.org/article_90313.html https://doi.org/10.31557/APJCP.2022.23.10.3247 |
| spellingShingle | Veterinary Sciences Suardana, I Wayan Swacita, Ida Bagus Ngurah Pinatih, Komang Januartha Putra Suharsono, Hamong Widiasih, Dyah Ayu Assessment of Cell Cycle and Induction of Apoptosis by Shiga-like Toxin Produced by Escherichia coli O157:H7 in T47D Breast Cancer Cells Using Flow Cytometry |
| title | Assessment of Cell Cycle and Induction of Apoptosis by
Shiga-like Toxin Produced by Escherichia coli O157:H7 in
T47D Breast Cancer Cells Using Flow Cytometry |
| title_full | Assessment of Cell Cycle and Induction of Apoptosis by
Shiga-like Toxin Produced by Escherichia coli O157:H7 in
T47D Breast Cancer Cells Using Flow Cytometry |
| title_fullStr | Assessment of Cell Cycle and Induction of Apoptosis by
Shiga-like Toxin Produced by Escherichia coli O157:H7 in
T47D Breast Cancer Cells Using Flow Cytometry |
| title_full_unstemmed | Assessment of Cell Cycle and Induction of Apoptosis by
Shiga-like Toxin Produced by Escherichia coli O157:H7 in
T47D Breast Cancer Cells Using Flow Cytometry |
| title_short | Assessment of Cell Cycle and Induction of Apoptosis by
Shiga-like Toxin Produced by Escherichia coli O157:H7 in
T47D Breast Cancer Cells Using Flow Cytometry |
| title_sort | assessment of cell cycle and induction of apoptosis by shiga like toxin produced by escherichia coli o157 h7 in t47d breast cancer cells using flow cytometry |
| topic | Veterinary Sciences |
| url | https://repository.ugm.ac.id/281800/1/Widiasih_KH.pdf |
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