The Investigation Of Stability Of KLF4 Protein Extracted From Stem Cells Using Different Spectroscopic Techniques Due ToCancer Prevention

The Kruppel-like factor (KLFs) family of gene regulatory proteins are transcription factorsimplicated in the regulation of a wide range of cellular processes, including proliferation,apoptosis, differentiation, inflammation, migration, and tumor formation. Kruppel-like factor 4(KLF4) is highly expressed in more than 70% of breast cancers and functions as an oncogene.However, an exact mechanism by which KLF4 enhances tumorigenesis of breast cancer remainsunknown. Down regulation of KLF4 in colon adenomas, gastric cancer, intestinal adenomas,esophageal cancer, prostate cancer, and lung cancer may contribute to cellular hyperproliferationand malignant transformation, which is consistent with its role in cell cycle arrest and growthinhibition. However, high levels of KLF4 expression are also reported in primary breast ductalcarcinoma and oral squamous carcinoma. So our findings have indeed pointed out the importance ofthe evaluation of conditions which can cause the instability of this protein due to prevent theprogression of cancers related to KLF4. To evaluate this process, we used some spectroscopictechniques such as Circular dichroism (CD), Three-Dimentional (3D) Fluorescence Spectroscopy2ml of %0.01 KLF4 in KH2PO4 buffer in various amounts of pH and ionic strengths was added and theranges were measured. The 3D spectra and contour maps of the KLF4 appears some peaks that eachone shows some features of the protein structure; The results shows that in all amounts of pH, theranges of Δλ in both peaks 1 and 2 are the same, indicates that transition of protein chromophorshad no modification. The most measure of fluorescence intensity of peak 1 and 2 was related toethanol-distilled water 50% solution that shows structural changes in protein that causeschromophor transitions to more polar environments and Tyr and Trp residues are involved tosolvent. But the most reduction of fluorescence intensity of peaks 1 and 2 was related to ethanoldistilledwater 100% that implicates chromophor transitions to more homophobic environments Inthis study, secondary structural changes of protein in different environments had been studied byCD, which show the dependence of protein to pH, which at pH=7.4 protein maintains it’s secondarystructure better and from pH=7.4 to the higher and lower pH, secondary structure will experiencemany changes.