Figure 8 Efficient P53 knockdown in cancer cells increases cellular sensitivity to TAI-1. (A) A549 and HCT116 cells which carry wild-type P53 were transfected with control siRNA (siControl) or P53 siRNA (siP53) for 24 hours and treated with TAI-1 (starting dose 100 μM, 3x serial dilution), incubated for 48 hours and analyzed for viability with MTS. Cellular sensitivity is expressed in GI50 (nM) and RNA from transfected cells were analyzed
for P53 RNA level by quantitative real time PCR. SiP53 reduced GI50s of compound in cells. (B) Selected cell lines which carry wild type P53 (A549, HCT116, ZR-75-1, U2OS) or mutated P53 (HeLa, as control) were transfected with siP53, treated with TAI-1 and analyzed for viability with MTS. Cellular sensitivity is expressed as% growth inhibition and cell lysates from transfected cells were collected and P53 protein levels click here determined by western https://www.selleckchem.com/products/netarsudil-ar-13324.html blotting. Differential Hec1 expression in clinical cancer
subtypes Genome-wide expression profile analysis has shown that Hec1 is upregulated in lung, colorectal, liver, breast, and brain tumors and that Hec1 expression correlates with tumor grade and prognosis [4, 9]. To determine whether HEC1 expression varies between cancer subtypes from the same tissue or organ, the gene expression data of NDC80 (HEC1) between adenocarcinoma and squamous carcinoma was studied for lung cancer. As shown in Figure 9A, NDC80 expression is significantly higher in squamous cell carcinoma of lung than adenocarcinoma in all three independent datasets. One way hierarchical cluster analysis consistently showed that NDC80, NEK2, NUF2 and SPC25 were reproducibly clustered together in three different gene expression datasets (Figure 9B). All these four genes showed higher expression in squamous cell carcinoma of lung.
The results indicate that different subtypes of lung cancer could respond differently to the treatment of Hec1 inhibitor. The predictability of response to Hec1-targeted treatment according to Hec1 associated gene expression remains to be further studied; however, our results suggest Cell press such consideration for HEC1 or related gene expression may be an important factor in the design of personalized Hec1-targets treatment of cancers. Figure 9 Differential expression of NDC80 (Hec1) and genes associated with NDC80 between subtypes of non-small cell lung cancer. (A) NDC80 (Hec1) (Affymetrix Probeset ID 204162_at) expression between adenocarcinoma and squamous cell carcinoma of lung in three different independent datasets (GSE8894, GSE3141 and GSE37745). The unit of Y axis is logarithm of expression intensity to the base 2. ANOVA was used to compare these two subtypes of NSCLC.