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 comprising a receptor tyrosine kinase targeted agent for treating a tumor in a patient comprising:

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers capable of amplifying 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 agent by comparing the differential EGFR expression level of step (f) and a threshold level for differential EGFR gene expression.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers capable of amplifying 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 of step (f) and a threshold level for differential HER2-neu gene expression.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor sample and non-malignant sample;
• (d) subjecting the mRNA to amplification using a pair of oligonucleotide primers capable of amplifying a region of the EGFR gene, and a pair of oligonucleotide primers capable of amplifying 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 of step (f) and a threshold level for differential HER2-neu gene expression, and comparing the differential EGFR expression level of step (f) and a threshold level for differential EGFR gene expression.

A method for determining a chemotherapeutic regimen comprising a 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 tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the HER2-neu tumor tissue sample and HER2-neu non-malignant tissue sample or determining the amount of EGFR mRNA in the EGFR tumor tissue sample and EGFR non-malignant tissue sample;
• (e) obtaining a differential HER2-neu expression level or obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression of about 1.8, or comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression of about 1.8.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample or determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential HER2-neu epression level or obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression of about 1.8, or comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression of about 1.8.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression of about 1.8.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential HER2-neu expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression of about 1.8.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample and determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant amplified sample;
• (e) obtaining a differential HER2-neu expression level and obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression of about 1.8, and comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression of about 1.8.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample or determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential HER2-neu epression level or obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression, or comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant tissue sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample;
• (e) obtaining a differential HER2-neu expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression.

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

• (a) obtaining a tissue sample of the tumor;
• (b) obtaining a matching non-malignant tissue sample;
• (c) isolating mRNA from the tumor tissue sample and non-malignant sample;
• wherein said tissue samples are fixed or fixed and paraffin embedded prior to said mRNA isolation;
• (d) determining the amount of HER2-neu mRNA in the tumor tissue sample and non-malignant tissue sample and determining the amount of EGFR mRNA in the tumor tissue sample and non-malignant amplified sample;
• (e) obtaining a differential HER2-neu expression level and obtaining a differential EGFR expression level;
• and
• (f) determining a chemotherapeutic regimen comprising a receptor tyrosine kinase targeted agent by comparing the differential HER2-neu expression level of step (e) and a threshold level for differential HER2-neu gene expression, and comparing the differential EGFR expression level of step (e) and a threshold level for differential EGFR gene expression.