Gene expression in clinical samples data from databases NDC80 gene expression data in non small cell lung cancer were retrieved from publicly available database. Gene expression intensities were normalized with quantile normalization. NDC80 expression between adenocarcinoma and squamous car cinoma was compared for all three different datasets. Eight genes known to associate with NDC80 were iden tified. One way hierarchical clustering analysis for adenocarcinoma and squamous carcinoma of NSCLC was conducted by using R package software. Results Hec1 inhibitor TAI 1 is highly potent with a wide anti cancer spectrum The initial small molecule hits identified by Drs. Chen in Dr. WH Lees laboratory, INH1 and INH2, had micro molar potency on cancer cell lines.

Through medicinal chemical efforts to modify the hit structure, we have significantly improved the potency of the Hec1 targeted compound selleck chemical to low nanomolar level. The new compound, TAI 1, has a GI50 of 13. 48 nM, which is close to 1000 times improvement in potency compared to INH1. To characterize the potency of the new compound, TAI 1, a series of cancer cell lines were tested. The screen includes 31 cancer cell lines, is comprise of 12 cell lines from the NCI 60 panel, and includes breast cancer, leukemia, liver, lung, colon cancer, cervical cancer, prostate cancer and bone cancer with various cellular characteristics. Growth inhibition was quantitated with established MTS assay. As summarized in Table 1, TAI 1 inhibits cellular growth at nM levels for the majority of cancer cell lines screened.

To determine the activity of TAI 1 in multidrug resist ant cell lines, established MDR cell lines were tested. MES SA Dx5 and NCI ADR RES are resistant to doxorubicin and paclitaxel, FR 180204 molecular weight while K562R cells are resist ant to imatinib. TAI 1 was active in these cell lines showing nM GI50. TAI 1 targets the Hec1 Nek2 pathway and induces apoptotic cell death To confirm the mechanism of action of TAI 1, we used established methods to evaluate the interaction of Hec1 and Nek2 and the consequences of disruption of inter action of the proteins. Co immunoprecipitation study shows that TAI 1 disrupted the binding of Nek2 to Hec1 in TAI 1 treated cells. Disruption of Nek2 binding to Hec1 was shown to lead to degradation of Nek2, and this was also confirmed for TAI 1.

In addition, previous study also show that disruption of Hec1 Nek2 interaction leads to misaligned chromosomes. Treatment of cells with TAI 1 induced a time dependent increase in the proportion of cells with chromosomal misalignment in cells. These results are consistent with the phenotypic consequences of the original hit compound INH1 and show that TAI 1 targets Hec1 Nek2 interactions. The cell death pathway was evaluated with apoptotic markers. Results show that TAI 1 induces cancer cell death through the induction of cleavage of apoptotic proteins Caspase 3 and PARP and degradation of anti apoptotic proteins MCL 1 and suggests that TAI 1 leads to activation of the apoptotic pathways.