Title

The Effect Of Paclitaxel And Cisplatin On A Variety Of Metastatic Mammary Heterogeneous Nuclear Ribonucleoprotein – E1 (HnRMP-E1) Knock Down Cell Lines

Author(s)

Dae Bigach

School Name

South Carolina Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Physiology and Health

Presentation Type

Mentored

Mentor

Mentor: Philip Howe, Molecular Biology Department, Medical University of South Carolina

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.

Start Date

4-11-2015 10:30 AM

End Date

4-11-2015 10:45 AM

COinS
 
Apr 11th, 10:30 AM Apr 11th, 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.