Method of determining a chemotherapeutic regimen based on ERCC1 and TS expression
The present invention relates to prognostic methods which are useful in medicine, particularly cancer chemotherapy. The object of the invention to provide a method for assessing TS and/or ERCC1 expression levels in fixed or fixed and paraffin embedded tissues and prognosticate the probable resistance of a patient's tumor to treatment with 5-FU and oxaliplatin-based therapies by examination of the amount of TS and/or ERCC1 mRNA in a patient's tumor cells and comparing it to a predetermined threshold expression level for those genes. More specifically, the invention provides to oligonucleotide primer pairs ERCC1 and TS and methods comprising their use for detecting levels of ERCC1 and TS mRNA, respectively.
1. A method for determining a chemotherapeutic regimen comprising 5-Fluorouracil, oxaliplatin, or combination thereof for treating a tumor in a patient comprising:
(a) obtaining a tissue sample of the tumor and fixing the sample, to obtain a fixed tumor sample;
(b) isolating mRNA from the fixed tumor sample;
(c) subjecting the mRNA to amplification using a pair of oligonucleotide primers capable of amplifying a region of the Thymidylate Synthase (TS) gene having SEQ ID NO: 11, to obtain an amplified sample, wherein the oligonucleotide primers consist of the oligonucleotide primer pair SEQ ID NO: 3 and SEQ ID No: 4;
(d) determining the amount of TS mRNA in the amplified sample;
(e) comparing the amount of TS mRNA from step (d) to an amount of mRNA of an internal control gene to determine the difference between the amplified sample and internal control gene; and
(g) determining a chemotherapeutic regime comprising 5-flurouracil, oxaliplatin, or combination thereof based on the amount of TS mRNA in the amplified sample and the threshold level for TS gene expression.
2. The method of claim 1 wherein the tumor is a colorectal adenocarcinoma tumor.
3. The method of claim 1 , wherein, the threshold level of TS gene expression is about 7.5 times an internal control gene expression level.
4. The method of claim 1 wherein the internal control gene is β-actin.