Project Title

Expression of MKNK2 Isoforms in Cancer Cell Lines

Academic department under which the project should be listed

CSM - Chemistry and Biochemistry

Research Mentor Name

Rajnish Singh

Abstract (300 words maximum)

MKNK2 is a protein that has 2 isoforms (MKNK2 long(a), and MKNK2 short(b)). They phosphorylate and activate translation initiation factor eIF4E and have been implicated in some cancers. MKNK2a specifically is found to be downregulated in breast, lung, and colon tumors. There are cases where the MKNK2a to MKNK2b ratio changes in normal cell lines versus cancerous cell lines, such as breast tissue where there is found to be a lower percentage of 1:1 ratio in noncancerous cell lines versus cancerous cell lines. This research project was conducted as part of a Course-based Undergraduate Research Experience (CURE) within the CHEM 3512 lab course. The goal of this project is to determine if the two isoforms are expressed in cancer cell lines using western blotting. His-tagged MKNK2 isoforms were purified and the isoform specific antibodies were tested on the purified proteins. The antibodies are able to differentiate between the two isoforms and will be used to probe cellular lysates prepared from BHK-2 (Baby Hamster Kidney-21), Hep G2 (hepatoblastoma-derived) and U-2OS (Osteosarcoma) cell lines. Data from this study will provide useful information on endogenous expression of MKNK2 isoforms in cancer and help provide preliminary contributions for further studies.

Disciplines

Biochemistry | Biochemistry, Biophysics, and Structural Biology | Laboratory and Basic Science Research | Life Sciences | Molecular Biology

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Expression of MKNK2 Isoforms in Cancer Cell Lines

MKNK2 is a protein that has 2 isoforms (MKNK2 long(a), and MKNK2 short(b)). They phosphorylate and activate translation initiation factor eIF4E and have been implicated in some cancers. MKNK2a specifically is found to be downregulated in breast, lung, and colon tumors. There are cases where the MKNK2a to MKNK2b ratio changes in normal cell lines versus cancerous cell lines, such as breast tissue where there is found to be a lower percentage of 1:1 ratio in noncancerous cell lines versus cancerous cell lines. This research project was conducted as part of a Course-based Undergraduate Research Experience (CURE) within the CHEM 3512 lab course. The goal of this project is to determine if the two isoforms are expressed in cancer cell lines using western blotting. His-tagged MKNK2 isoforms were purified and the isoform specific antibodies were tested on the purified proteins. The antibodies are able to differentiate between the two isoforms and will be used to probe cellular lysates prepared from BHK-2 (Baby Hamster Kidney-21), Hep G2 (hepatoblastoma-derived) and U-2OS (Osteosarcoma) cell lines. Data from this study will provide useful information on endogenous expression of MKNK2 isoforms in cancer and help provide preliminary contributions for further studies.