Application of siRNA technology to examine roles of - Research

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					                                                Mentor Application
                                SCHOOL OF MEDICINE

  Please submit completed applications to Leslie Skelton in the Office of Research (U020) or electronically at by January 5, 2009. If you would like to have your application available to
           students before the winter holiday, please submit it no later than December 15, 2008.

Project Title: Application of siRNA technology to examine roles of Phospholipase C isoforms in the migration of HCT-15

Mentor Name: Ujjwal K. Rout
      Phone number: (601) 815 5193
      Department: Surgery

Number of Students Previously Supervised: 13
Supervised Student Productivity:
    Number of Student Papers: One under preparation
    Number of Student Abstracts: 8
    Number of Student Presentations: 8

Funding Source for Work: Yes

Would you be able to support a summer student’s salary without the Dean’s Grant? No

IRB Approval (Clinical Research) Obtained: N/A

IACUC Approval (Research with Animals) Obtained: N/A
Briefly describe your project so the student can clearly understand the work that will be undertaken and the rationale for doing the research. Do not exceed
these two pages. You should have the laboratory or clinical resources to support the proposed work. The project should have a reasonable chance of producing
tangible results during the 10-week length of the fellowship.

Statement of Problem and Background:

         Colorectal cancer (CRC) is one of the most common malignancies and the second leading cause of
          death from cancer in Europe and North America. It is responsible for approximately one million new
          cases and half a million deaths per year worldwide.
         Metastasis of colorectal cancer cells (their propagation and colonization of distant organs including liver
          and brain) may continue even after the surgical removal of the cancerous tissue and is the leading
          cause of death due to colon cancer.
         The process of metastasis is similar to the embryonic cell migration and wound healing. Molecules
          responsible for the cancer cell migration include integrins, matrix-degrading enzymes, and cell-to-cell
          adhesion and communication molecules.
         Previous studies under my mentorship (summer student research, 2008) demonstrate that beta1
          integrin subunit is involved in the migration of human colon cancer cell line (HCT-15). However, our
          attempts to examine the involvement of phospholipase C gamma 1 (PLC) isoform, a molecule that is
          known to be involved in the migration of cells, did not alter the migration of these cells. This is a
          surprising and interesting observation because it indicates that human colon cancer cells may use
          different isoform of PLCor possibly the path of integrin signaling not involving PLC.
         Due to non-availability of commercially available monoclonal antibodies against different PLC isoforms,
       we propose to use siRNA technology to identify isoform/s of PLC- (PLC-and PLC-2) that may be
       involved in the migration of HCT-15 cells.

Methods and Data Analysis:

      Expression of different isoforms of PLC in the HCT cells will be examined using RT-PCR techniques.
      A previously published method by the mentor for studying cell migration will be used for studying the
       migration of HCT-15 cells in culture using microscopy and Metamorph software.
      siRNA and appropriate control molecules specific to different PLC isoforms will be examined on the
       migration of colon cancer cell line (HCT-15).

Anticipated Outcome(s):

      Because siRNA molecules are synthesized based on the cDNA sequence available from the nucleic
       acid data base and are highly specific in reducing the expression of target molecule of interest inside a
       cell, we anticipate to determine whether any of the PLC- isoform/s is required for the migration of HCT-
       15 cells.
      It is anticipated that PLC-isofom/s other than PLC-1, (PLC-2) may be involved in the integrin receptor
       mediated signaling and migration of HCT-15 cells.

Significance of the Work:

      Several clinical trials are undergoing to test the efficacy of integrin receptor signaling in the migration of
       cancer cells. Because colon cancer is one of leading cause of death due to cancer, identification of
       specific PLC- isoform may allow designing effective drugs to inhibit the metastasis of colon cancer.
      Therefore, studies suggested in this application would allow the investigators to determine important
       unknowns that have significant clinical implications in the treatment strategy of colon cancers.

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