Regulation Cancer Proliferation using siRNA Technology

Abstract

There has been a great attention towards the immunotherapeutic strategies for the treatment of cancer due to their potential capability of eradicating metastases and cancerous cells by means of employing the host immune system. While alternatively, the therapeutics based on the RNA having the capability of hushing the appearance of particular targets are presently being investigated clinically for several syndromes counting in cancer. Due to the versatility of the methods for evading tumor from the host immune system, varying molecules are capable of being targeted by the technology of RNAi for the purpose of enhancing the invulnerable response against cancers. This technology has been employed for silencing particular targets in cancerous cells and also the immune cells in the lines of cancer cells, critical trials and animal models. Furthermore, siRNAs are able of stimulating the inherent immune responses by means of introduction of Toll-like receptors. Despite the fact that the present clinical examinations of the employment of siRNA technology are considerably less in number, yet it can be foreseen that this technology in prospect would be broadly employed in the treatment of cancer.

Table of Contents

I. INTRODUCTION4

II. EXPERIMENTAL DESIGN5

Comparison of the OTUB 1 expression level in normal and cancer cell5

Checking the effect of siRNA in knocking down OTUB 1 gene5

Effect on siRNA on cell proliferation of H3586

Proliferation pattern of cancer cell when siRNA coexists with anti-cancer material doxorubicin6

III.MATERIALS AND METHODS6

Cell Culture6

Western blotting7

Proliferation Assay7

Transfection of siRNA8

Treatment of Doxorubicin8

IV.RESULTS8

Comparison of the OTUB 1 expression level in normal and cancer cell8

Checking the effect of siRNA in knocking down OTUB 1 gene9

Effect on siRNA on cell proliferation of H35812

Proliferation pattern of cancer cell when siRNA co-exists with anti-cancer material doxorubicin14

V.DISCUSSIONS16

VI. REFERENCES18

Regulation Cancer Proliferation using siRNA Technology

I. Introduction

Cancer cells have a different element in them that allows them to proliferate uncontrollably and therefore, experiment will be conducted in order to find out what gene is causing these cells to multiply greatly. When one specific gene is causing this endless growth, that gene needs to be suppressed in order to see if that affects the proliferation rate of these cancer cells. This phenomenon of suppressing a specific gene is called knock down.

Also, in order to knock down the gene that appears the most, siRNA technology is used. siRNA refers to a small interfering or short interfering RNA. This RNA requires transfection of cells using a lipid-based transfection reagent. This lipid-based transfection causes the nucleic acids to enter into the mammalian cell, in this case, the lung cancer cell, easily. This technology is also useful for a transient knock-down, which refers to the change in gene expression without the modification of a chromosomal DNA (Santacruz Biotechnology Incorporation).

Furthermore, we have to consider the possibility of a proliferation that is caused by a protein that is over-produced. If that occurs, the protein needs to be determined in order to stop it from being over-produced.

The two different cells that will be used are WI-38 and H358. WI-38 is a human diploid normal cell from the ...