Valproic Acid Negatively Regulated CD44 and BMI1 Expression in Stem†like Cancer Cells

Cancer therapy is still very challenging since current modalities are not effective enough toeradicate chemoradiotherapy resistance and metastatic cancer cells. Valproic acid (VPA) is ashort chain fatty acid with histone deacetylase-inhibiting effects that has been examined asan anticancer agent in several clinical trials. However, little is known about VPA effects onunique properties of cancer stem cell (CSCs). Therefore, the aim of present study was toinvestigate VPA effects on the expression of BMI1 and CD44, which have been introducedas CSC markers in gastrointestinal malignancies. In present study, KYSE30 cells, which areesophageal stem-like cancer cells positive for BMI1 and CD44, were treated with 2.5 and 5mM VPA for 48 and 72h. Then, the total cellular RNA was extracted, and cDNAs weresynthesized by M-MuLV reverse transcriptase. For quantitative RT-PCR, SYBR green mastermix was used and PCR efficiencies were calculated for both BMI1 and CD44 primers. Tonote, normalized values were plotted as relative fold change over untreated cells, and foldchange expression was calculated as 2(-ΠΠCT). Quantitative RT-PCR results indicated thatVPA significantly downregulated the expression of CSC markers in KYSE30 cells.Interestingly, in comparison with 5 mM VPA, treating cells with 2.5 mM VPA decreased theexpression of BMI1 and CD44 to lower levels; after 48 and 72h, BMI1expression wascalculated as 0.11 ± 0.02 and 0.08 ± 0.01, respectively, and CD44 expression decreasedto 0.4 ± 0.03 and 0.25 ± 0.03, respectively. KYSE30 cells present a suitable model forstudying effects of anticancer agents on stem-like cancer cells. Our results indicated that VPAhas negative regulatory effects on the expression of CSC markers. However, more researchis needed to understand whether VPA has similar effects on other CSC molecules, such asALDH1, EpCAM and LGR5.

کلیدواژه ها:

Valproic Acid ، Stem-like Cancer Cells ، BMI1 ، CD44 ، Quantitative RT-PCR