The Effect Of Paclitaxel And Cisplatin On A Variety Of Metastatic Mammary Heterogeneous Nuclear Ribonucleoprotein – E1 (HnRMP-E1) Knock Down Cell Lines
School Name
South Carolina Governor's School for Science and Mathematics
Grade Level
12th Grade
Presentation Topic
Physiology and Health
Presentation Type
Mentored
Abstract
Breast cancer is the most commonly diagnosed cancer in women (9). One protein Heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) when absent allows epithelial mesenchymal transition to occur in primary tumors. This protein was knocked down in a variety of Normal Murine Mammary Gland (NMuMG) cells so they would metastasize. The cells then underwent in vivo passage in mice to mimic a metastatic tumor returning in a patient. The resulting cell lines were named NMuMG, hnRNP E1 Knock Down (E1KD), mammary 1 passage (M1P), lung 1 passage (L1P), and lung 2 passage (L2P). The aim of this research was to test the drug resistance of these five cell lines. These cells lines were treated for four hours with two chemotherapeutic drugs at concentration gradients of 0uM to 1uM for Paclitaxel and 0uM to 20uM for Cisplatin. An MTT assay was performed to determine the viability of the cells. The results showed that the L2P cells had the highest rate of survival followed by the L1Ps, the E1KDs, the M1Ps, and lastly the NMuMGs, showing that the more metastatic cell lines were better able to overcome the drugs. To further characterize the cell lines, the five lines were then subjected to growth curves, RT-PCRs to look at mesenchymal markers, immunofluorescence microscopy to look at spindle growth, and flow cytometry to look at cell cycle patterns. Results indicate that the L2Ps lost contact inhibition and were able to overcome the G2/M checkpoint better than others in the cell line.
Recommended Citation
Bigach, Dae, "The Effect Of Paclitaxel And Cisplatin On A Variety Of Metastatic Mammary Heterogeneous Nuclear Ribonucleoprotein – E1 (HnRMP-E1) Knock Down Cell Lines" (2015). South Carolina Junior Academy of Science. 101.
https://scholarexchange.furman.edu/scjas/2015/all/101
Start Date
4-11-2015 10:30 AM
End Date
4-11-2015 10:45 AM
The Effect Of Paclitaxel And Cisplatin On A Variety Of Metastatic Mammary Heterogeneous Nuclear Ribonucleoprotein – E1 (HnRMP-E1) Knock Down Cell Lines
Breast cancer is the most commonly diagnosed cancer in women (9). One protein Heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) when absent allows epithelial mesenchymal transition to occur in primary tumors. This protein was knocked down in a variety of Normal Murine Mammary Gland (NMuMG) cells so they would metastasize. The cells then underwent in vivo passage in mice to mimic a metastatic tumor returning in a patient. The resulting cell lines were named NMuMG, hnRNP E1 Knock Down (E1KD), mammary 1 passage (M1P), lung 1 passage (L1P), and lung 2 passage (L2P). The aim of this research was to test the drug resistance of these five cell lines. These cells lines were treated for four hours with two chemotherapeutic drugs at concentration gradients of 0uM to 1uM for Paclitaxel and 0uM to 20uM for Cisplatin. An MTT assay was performed to determine the viability of the cells. The results showed that the L2P cells had the highest rate of survival followed by the L1Ps, the E1KDs, the M1Ps, and lastly the NMuMGs, showing that the more metastatic cell lines were better able to overcome the drugs. To further characterize the cell lines, the five lines were then subjected to growth curves, RT-PCRs to look at mesenchymal markers, immunofluorescence microscopy to look at spindle growth, and flow cytometry to look at cell cycle patterns. Results indicate that the L2Ps lost contact inhibition and were able to overcome the G2/M checkpoint better than others in the cell line.
Mentor
Mentor: Philip Howe, Molecular Biology Department, Medical University of South Carolina