PME-1 as a biomarker to predict and diagnose an increased risk of endometrial cancer and gene silencing of PME-1 to inhibit epithelial to mesenchymal transition
Disclosed are methods of attenuating activity of the PME-1 gene. siRNAs or shRNAs are used to target against PME-1, thereby reducing the PME-1 mRNA. It is disclosed that the siRNAs or shRNAs targeted against PME-1 attenuate the epithelial to mesenchymal transition, thereby inhibit endometrial cancer development. A kit containing siRNA or shRNA reagents for attenuating the PME-1 gene expression is also disclosed.
1. A method for determining whether a woman has endometrial cancer, comprising the steps of:
(a) obtaining an endometrial tissue from a woman suspected of having an increased risk of endometrial cancer;
(b) obtaining a normal endometrial tissue;
(c) treating said endometrial tissue obtained in step (a) with a lysis buffer to prepare a first lysate;
(d) treating said endometrial tissue in obtain step (b) with a lysis buffer to prepare a second lysate;
(e) performing a Western blot or an ELISA to quantify PME-1 protein expression level in said first lysate and PME-1 protein expression level in said second lysate;
(f) comparing said PME-1 protein expression level in said first lysate with said PME-1 protein expression level in said second lysate; and
(g) diagnosing said woman as having endometrial cancer, if there is an increased PME-1 protein expression level in said endometrial tissue obtained in step (a) relative to said normal endometrial tissue obtained in step (b),
wherein said Western blot or ELISA uses a goat, rabbit or mouse antibody.
2. A method of detecting PME-1 in a patient, comprising the steps of:
(a) obtaining an endometrial tissue from a woman suspected of having an increased risk of endometrial cancer;
(b) performing a Western blot or an ELISA to on the endometrial tissue to quantify PME-1 protein expression level in said tissue.
3. The method of claim 2 wherein said endometrial tissue obtained in step (a) is treated with a lysis buffer to prepare a lysate, and the Western Blot or ELISA is performed on the lysate.
4. The method of claim 3 , wherein said lysis buffer is a modified RIPA solution.
5. The method of claim 2 , wherein said step (b) is performed by a Western blot.
6. The method of claim 2 , wherein said step (b) is performed by an ELISA.
7. The method of claim 5 , wherein said Western blot is performed using an anti-PME-1 monoclonal antibody or polyclonal antibody.
8. The method of claim 6 , wherein said ELISA is performed using an anti-PME-1 monoclonal antibody or polyclonal antibody.
9. The method of claim 2 , wherein said endometrial tissue is grade II or grade III cancer tissue.
10. The method of claim 2 wherein said Western blot or ELISA uses a goat, rabbit or mouse antibody.