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The role of “mitotic slippage” in the origin of amoeboid metastatic cell in therapy resistant cancers

 

 

Funding: European Regional Development Fund (ERDF) “On Implementation of Activity 1.1.1.2 “Post-doctoral Research Aid” of the Specific Aid Objective 1.1.1 “To increase the research and innovative capacity of scientific institutions of Latvia and the ability to attract external financing, investing in human resources and infrastructure” of the Operational Programme “Growth and Employment”

Project Title: The role of “mitotic slippage” in the origin of amoeboid metastatic cell in therapy resistant cancers

Project No.: 1.1.1.2/VIAA/3/19/463

Period: 36 months (1 January 2020 – 31 December 2022)

Project costs: 133 806.00 EUR

Project implementer: Dr. biol. Kristīne Salmiņa

 

The mortality of cancer patients still remains high, mostly due to metastatic disease, where tumor cell migration is a critical mechanism in development of resistance to anticancer therapy.  The amoeboid motility is a very efficient mode of migration and metastatic growth of single tumor cells. The working hypothesis of this project suggests the role of “mitotic slippage” – reverse of aborted mitosis enhanced by therapy in formation of polyploid tumour cells undergoing the reprogramming to amoeboid state. Beside motility, amoeboid cancer cells display various activities cooperatively enabling survival and drug-resistant metastatic growth. Impact of the key amoebal cdc42-kinase on these activities will be characterised in mechanistic experiments. The work will be performed on breast cancer and melanoma cell lines, and include the patient material. Genotoxic treatments, cdc42 inhibition will be examined by immunofluorescence, cytometry, western blot, qRT-PCR, live-imaging and confocal microscopy. In addition, the clonogenicity, migration, invasion, and tumorigenicity tests will be applied.

Information published 02.01.2020.

 

Project progress

 1 January 2020 – 31 March 2020

At this stage of the project, the breast cancer MDA-MB 231 cell line was treated with the anticancer agent doxorubicin, and mitotic slippage, which is the source of giant cell formation, was observed. The cell cycle, polyploidization, growth curve and microscopic counts of mitotic slippage were determined. Activation of the meiotic genes MOS, DMC1, REC8 and SPO11 after cell treatment was observed. The signs of amoeboidisation in giant cells resulting from the application of anti-cancer therapy in comparison to control cells have been investigated. Work has been done to prepare and submit the publication.

Information published 31.03.2020. 

 

Project progress

 1 April 2020 – 30 June 2020

At this stage of the project, the human melanoma SK-MEL 28 cell line was treated with the anticancer agent Vemurafenib and the cell cycle, growth curve and expression of meiotic genes after optimization of cell treatment were determined. However, increased cell polyploidisation and mitotic slippage were not observed after treatment. A scientific article on the role of mitotic slippage and extranuclear DNA in cancer cell resistance has been published: doi: 10.3390/ijms21082779.

Information published 30.06.2020. 

 

Project progress

 1 July 2020 – 30 September 2020

At this stage of the project, the human melanoma SK-MEL 28 cell line was treated with the anticancer agent doxorubicine. Optimization of cell treatment and characterization of resistance model were performed. Cell cycle analysis, polyploidisation, growth curve and microscopic detection of mitotic slippage were carried out, as well as the expression of meiotic kinase MOS and CyclinB1 using immunofluorescence was studied. It was found that doxorubicine in this cell line, as in MDA-MB 231 cells, induces mitotic slippage and subsequent polyploidisation.

Information published 30.09.2020.

 

Project progress

1 October 2020 – 31 December 2020

At this stage of the project, samples were obtained for the analysis of the transcription profile at different time points after treatment of MDA-MB 231 and SK-MEL 28 cell lines with doxorubicin. Work has been done on the preparation and submission of the publication, as a result of which a scientific review article has been published https://doi.org/10.1016/j.semcancer.2020.12.009

Information published 30.12.2020. 

 

Project progress

 1 January 2021 – 31 March 2021

At this stage of the project, the clonogenicity, the expression of meiotic genes and senescence markers of SK-MEL28 cells after treatment with doxorubicin and vemurafenib were studied. The results of the project were also presented at the conference (MDPI: The 1st International Electronic Conference on Cancers: Exploiting Cancer Vulnerability by Targeting the DNA Damage Response) as a poster presentation 10.3390 / IECC2021-09214.

Information published 30.03.2021. 

 

Project progress

 1 April 2021 – 30 June 2021

At this stage of the project, RNA was isolated and purified from samples of MDA-MB 231 and SK-MEL 28 cell lines obtained before and at different time points after genotoxic treatment. Work has been done on the preparation and submission of the publication, as a result of which a scientific review article has been published https://doi.org/10.3390/cells10071582.

Information published 30.06.2021. 



Mājas lapas izstrādi finansēja ERAF 2.1.1.2. aktivitātes projekts Nr. 2010/0196/2DP/2.1.1.2.0/10/APIA/VIAA/004 "Latvijas biomedicīnas pētījumu integrācija Eiropas zinātnes telpā".