Method of determining epidermal growth factor receptor and HER2-neu gene expression and correlation of levels thereof with survival rates

A method for determining a chemotherapeutic regimen receptor tyrosine kinase targeted agent for treating a tumor in a patient comprising:

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a non-malignant tissue sample matching said tumor,
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the EGFR gene, to obtain an tumor amplified sample and a non-malignant amplified sample;
• (e) determining the amount of EGFR mRNA in the tumor amplified sample and non-malignant amplified sample;
• (f) obtaining a differential EGFR expression level, and
• (g) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted by comparing the differential EGFR expression level and the threshold level for EGFR gene expression.

A method for determining a chemotherapeutic regimen comprising receptor tyrosine kinase targeted agent for treating a tumor in a patient comprising:

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a non-malignant tissue sample matching said tumor;
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the HER2-neu gene, to obtain an tumor amplified sample and a non-malignant amplified sample,
• (e) determining the amount of HER2-neu mRNA in the tumor amplified sample and non-malignant amplified sample;
• (f) obtaining a differential HER2-neu expression level;
• and
• (g) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level and the threshold level for HER2-neu gene expression.

A method for determining a chemotherapeutic regimen comprising receptor tyrosine kinase targeted agent for treating a tumor in a patient comprising:

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a non-malignant tissue sample matching said tumor;
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the EGFR gene, and a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the HER2-neu gene, to obtain an EGFR tumor amplified sample and an EGFR non-malignant amplified sample, and a HER2-neu tumor amplified sample and a HER2-neu non-malignant amplified sample
• (e) determining the amount of HER2-neu mRNA in the HER2-neu tumor amplified sample and HER2-neu non-malignant amplified sample and determining the amount of EGFR mRNA in the EGFR tumor amplified sample and EGFR non-malignant amplified sample;
• (f) obtaining a differential HER2-neu expression level and obtaining a differential EGFR expression level;
• and
• (g) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level and the threshold level for HER2-neu gene expression, and comparing the differential EGFR expression level and the threshold level for EGFR gene expression.

A method for determining the level of EGFR expression in a fixed paraffin embedded tissue sample comprising;

• (a) deparaffinizing the tissue sample, to obtain a deparaffinized sample;
• (b) isolating mRNA from the deparaffinized sample in the presence of an effective amount of a chaotropic agent;
• (c) subjecting the mRNA to amplification using a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the EGFR gene, to obtain an amplified sample,
• (d) determining the quantitiy of EGFR mRNA relative to the quantity of mRNA of an internal control gene.

A method for determining the level of HER2-neu expression in a fixed paraffin embedded tissue sample comprising;

• (a) deparaffinizing the tissue sample, to obtain a deparaffinized sample;
• (b) isolating mRNA from the deparaffinized sample in the presence of an effective amount of a chaotropic agent;
• (c) subjecting the mRNA to amplification using a pair of oligonucleotide primers that hybridize under stringent conditions to a region of the HER2-neu gene, to obtain an amplified sample;
• (d) determining the quantity of HER2-neu mRNA relative to the quantity of mRNA of an internal control gene.