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Prostate carcinogenesis predictor

a prostate cancer and predictor technology, applied in the direction of instruments, transferases, drug compositions, etc., can solve the problem of achieve the effect of reducing cell proliferation, reducing cell cycle, and maintaining static control of prostate cancer

Inactive Publication Date: 2009-05-21
UNIV OF SOUTH FLORIDA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0003]As described herein, the invention consists of a means and product to detect protein kinase C-iota (PKC-ι) levels in prostate glands to identify prostate carcinogenesis. Prostate cancers are highly lethal tumors and are the second most prevalent type of cancer (following skin cancer) among men in the United States. Approximately one third of men diagnosed with cancer each year will have prostrate cancer. Despite significant educational efforts, improved diagnostic techniques, and rigorous therapies, prostate cancer control remains static. To address this health issue, the presence of PKC-ι in normal prostate tissue and prostate cancer tissue was investigated, to establish if PKC-ι could be used as a predictor of carcinogenesis. Western blots probing for PKC-ι were performed on 7 normal prostate biopsies, 2 prostate intraepithelial neoplasia and 14 prostate cancers. Results demonstrated minimal or no detection of PKC-ι the in normal prostate tissue. In comparison, PKC-ι was robustly present in prostate intraepithelial neoplasia (PIN). Similarly, PKC-ι was present in all prostate cancer biopsies. The increase in PKC-ι in prostate cancer biopsies was 100 fold compared to normal prost

Problems solved by technology

Despite significant educational efforts, improved diagnostic techniques, and rigorous therapies, prostate cancer control remains static.

Method used

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Examples

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example 1

PKC-ι is Overexpressed in Malignant or Neoplastic Prostate Tissues

[0359]The relationship between the absence of PKC-ι in BPH tissue and its robust presence in malignant prostate tissue is summarized in Table 1. Western blots probing for PKC-ι in 6 BPH biopsies, 3 PIN, and 7 malignant tumors revealed absence of PKC-ι in BPH (Table 1). By comparison, PKC-ι was robustly present in two PIN and absent in one. PKC-ι was abundant in all malignant prostate tissue. Western blots corresponding to some of the data present in Table 1 are shown in FIGS. 1A and 1B. PKC-α and PKC-ι were identified in Western blots by bands with molecular weights of 80 kD and 67 kD, respectively which corresponded to the immunoreactive signal obtained from U-373MG glioma cells which contain PKC-α and PKC-ι (data not shown). Western blot controls for PKC-α did not show a pattern of expression specific to BPH, PIN or malignant prostate tumors. Control β-actin Western blots showed β-actin immunoreactive bands at a mol...

example 2

Immunoblot Analysis of PKCs in siRNA Treated Prostate Cells

[0362]Immunoblotting of PKC-ι expression shows no effect in control siRNA (See FIG. 2 A-C). After counting the cells (from FIG. 3), the same populations of treated cells were immunoblot for PKC-ι, as described in “Materials and Methods.” A protein concentration of 15 μg was loaded on each lane for each time point and separated by SDS-PAGE and Western blotted with anti-PKC-ι mouse monoclonal or anti-PKC-ζ rabbit polyclonal antibody. Immunoblotting for PKC-ι in PKC-ι siRNA treated RWPE-1 cells showed that expression decreased by 90-92%. Absorbance densitometry (FIGS. 2A and 2D) shows the average of three independent PKC-ι immunoblot for both RWPE-1 cell treatments (p=0.013, 0.005, 0.029). To show specificity, we immunoblotted for PKC-ζ isoforms which is 98% identical to PKC-ι (FIG. 2A, second row).

[0363]There was no or very little PKC-ι present in PKC-ι siRNA treated cells compared to control siRNA. Furthermore, PKC-ι siRNA ha...

example 3

PKC-ι siRNA Treatment Decreases Prostate Cell Viability and Inhibits Cell Cycle Progression

[0365]To provide additional evidence that PKC-ι is required for cell proliferation in RWPE-1 cells, PKC-ι was temporarily inhibited using PKC-ι siRNA. Subconfluent RWPE-1 (A), LNCaP (B) and DU-145 (C) cells (2.5×105) were treated with PKC-ι siRNA (50 nM for RWPE-1, 100 nM for LNCaP and 150 μM for DU-145) complex from 24-72 h as described in “Materials and Methods.” At the indicated time, the viable cells were counted using trypan blue exclusion assay and a hematocyometry. Three independent experiments were performed and the mean of the viable cells and SD were plotted. After titrating PKC-ι siRNA concentrations from 50-100 nM, we used an optimal concentration of PKC-ι siRNA (50 nM) and control siRNA to suppress PKC-ι expression. It was shown that there is a decrease in cell proliferation in PKC-ι siRNA treated cells compared to control siRNA (FIG. 3A-C). Their standard values were as follow: R...

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Abstract

A method of detecting prostate tumorigenesis in a subject, the method including the steps of (a) obtaining a sample from the prostate of the human subject, (b) detecting quantitatively or semi-quantitatively in the sample a level of expression for PKC-ι and (c) comparing the expression level in (b) to a level of expression in a normal control, wherein overexpression of PKC-ι, with respect to the control, indicates the presence of prostate cancer in the subject. The present invention is based upon the discovery that PKC-ι levels are elevated during prostate tumorigenesis. Furthermore, the proliferation rate of the tumor correlates with the level of PKC-ι. The invention also provides methods of treating prostate cancer by administering to the subject a compound that inhibits the expression of PKC-ι. The compound can be a small interfering RNA (siRNA) molecule.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application claims the benefit of U.S. provisional application Ser. No. 60 / 980,611, filed Oct. 17, 2007, which is herein incorporated by reference in its entirety.FIELD OF INVENTION[0002]This invention relates to detection and treatment of prostate cancer. More specifically, the invention relates to use of protein kinase C-iota (PKC-ι) in the detection of prostate cancer and methods of inhibiting the expression and activity of protein kinase C-iota as a treatment for prostate cancer.SUMMARY OF INVENTION[0003]As described herein, the invention consists of a means and product to detect protein kinase C-iota (PKC-ι) levels in prostate glands to identify prostate carcinogenesis. Prostate cancers are highly lethal tumors and are the second most prevalent type of cancer (following skin cancer) among men in the United States. Approximately one third of men diagnosed with cancer each year will have prostrate cancer. Despite significant educat...

Claims

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Application Information

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IPC IPC(8): A61K9/127G01N33/53G01N33/535A61P35/00C12Q1/68A61K31/7105
CPCA61K9/127A61K31/7105C12N15/1137C12N2310/14Y10T436/25G01N33/57434G01N2800/56A61K31/675A61K31/713C12Y207/11013A61P35/00
Inventor ACEVEDO-DUNCAN, MILDREDWIN, HLA Y.SALUP, RAOUL
Owner UNIV OF SOUTH FLORIDA
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