Using comparative genome hybridization array analysis you have identified two ge

Question

Using comparative genome hybridization array analysis you have identified two genes (1 and 2) that are deleted in liver cancer cells. You wish to test whether either if these two genes can act as a tumor suppressor. For this, you add siRNA targeting each gene to a non-tumorigenic liver cell line. You assay foci formation and obtain the following results. Is either Gene 1 or Gene 2 functioning as a tumor suppressor? Explain your answer. How could you determine if Gene 1 and Gene 2 are reduced in expression in liver cancer cells relative to healthy cells. Be specific in your answer, do not just name the technique. Do you think that this type of liver cancer could possibly be treated by gene therapy? Explain your answer. Give one example of a tumor suppressor and briefly state how it functions to prevent tumor growth. (You may continue your answer on the back of this page if you need more space.)

Explanation / Answer

(a). In siRNA targeting Gene 2, No foci formation was detected. So Gene 2 may act as a Suppresser.

(b) siRNA was made to target different regions of Liver to affect different genes. And expression of each gene was monitored using quantitative real time RT-PCR. Effect of siRNA was tested on cacerous liver cells by observations of all the cell processes like invasion, proliferation and foci formation, growth by using Soft agar assay, apoptosis by annexing binding , and cell cycle arrest etc.

A decrease in focus forming potential, proliferation and migration capabilities indicates reduced expression.

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