Alternative splice variant patterns of human telomerase reverse transcriptase (hTERT) in thyroid tumors to distinguish benign from malignant
This invention relates, e.g., to a method for determining if a thyroid tumor in a subject is malignant, comprising determining in a sample from the subject the amount of TERT (telomerase reverse transcriptase) mRNA which lacks the β sequence and the amount of TERT mRNA in the sample which comprises the β sequence, wherein a preponderance (e.g., at least about 55%) of TERT mRNA in the sample which comprises the β sequence indicates that the tumor is malignant, and wherein a preponderance of TERT mRNA which lacks the β sequence indicates that the tumor is not malignant.
1. A method for determining if a thyroid tumor in a human subject is malignant, comprising the steps of:
(a) obtaining a sample from the subject;
(b) determining in the sample from the subject the amount of human TERT (telomerase reverse transcriptase (hTERT)) mRNA that lacks the β sequence and the amount of hTERT mRNA in the sample that comprises the β sequence using quantitative real time polymerase chain reaction (PCR), wherein the quantitative real time PCR utilizes nested primers and wherein the nested primers comprise SEQ ID NO:9 and SEQ ID NO:10; and
(c) identifying the tumor as malignant if the hTERT mRNA in the sample that comprises the β sequence comprises at least 55% of hTERT mRNA in the sample and identifying the tumor as not malignant if the hTERT mRNA that lacks the β sequence comprises at least 55% of hTERT mRNA in the sample.
2. The method of claim 1 , wherein the β sequence is the 182 base pair sequence represented by SEQ ID NO:3.
3. A method for selecting a treatment method for a subject having a thyroid tumor, comprising determining by a method of claim 1 whether the tumor is malignant and, if the tumor is determined to be malignant, deciding to perform a total thyroidectomy on the subject, but if a tumor is determined not to be malignant, deciding not to perform a total thyroidectomy on the subject.
4. The method of claim 1 , wherein the method is carried out both before or at approximately the same time as, and after, the administration of a treatment for thyroid cancer, and which is a method for determining the effectiveness of the treatment.
5. A method for treating a subject having a thyroid tumor, comprising determining by a method of claim 1 whether the tumor is malignant and, if the tumor is malignant, treating the subject aggressively for thyroid cancer, and if the tumor is determined not to be malignant, not treating the subject aggressively for thyroid cancer.
6. The method of claim 1 , wherein in the identification step (c) identifying the tumor as malignant if the hTERT mRNA in the sample that comprises the β sequence comprises at least 59% of hTERT mRNA in the sample and identifying the tumor as not malignant if the hTERT mRNA that lacks the β sequence comprises at least 59% of hTERT mRNA in the sample.
7. A method for determining if a thyroid tumor in a human subject is malignant, comprising the steps of:
(a) obtaining a sample from the subject;
(b) determining in the sample from the subject the amount of human TERT (telomerase reverse transcriptase (hTERT)) mRNA that lacks the β sequence and the amount of hTERT mRNA in the sample that comprises the β sequence using quantitative real time polymerase chain reaction (PCR), wherein the quantitative real time PCR utilizes nested primers and wherein the nested primers comprise SEQ ID NO:9 and SEQ ID NO:10; and
(c) identifying the tumor as malignant if the ratio of hTERT mRNA in the sample that comprises the β sequence compared to the total amount of hTERT mRNA in the sample is at least 0.55 and identifying the tumor as not malignant if the ratio of hTERT mRNA in the sample that lacks the β sequence compared to the total amount of hTERT mRNA in the sample is at least 0.55.
8. The method of claim 7 , wherein in the identification step (c) identifying the tumor as malignant if the ratio of hTERT mRNA in the sample that comprises the β sequence compared to the total amount of hTERT mRNA in the sample is at least 0.59 and identifying the tumor as not malignant if the ratio of hTERT mRNA in the sample that lacks the β sequence compared to the total amount of hTERT mRNA in the sample is at least 0.59.
9. A method for treating a human subject having a malignant thyroid tumor comprising the steps of:
(a) obtaining a sample from the subject;
(b) determining in the sample from the subject the amount of human hTERT mRNA that lacks the β sequence and the amount of hTERT mRNA in the sample that comprises the β sequence using quantitative real time polymerase chain reaction (PCR), wherein the quantitative real time PCR utilizes nested primers and wherein the nested primers comprise SEQ ID NO:9 and SEQ ID NO:10;
(c) performing a total thyroidectomy on the subject if the hTERT mRNA in the sample that comprises the β sequence comprises at least 55% of hTERT mRNA in the sample.
10. The method of claim 9 , wherein in the performing step (c) performing a total thyroidectomy on the subject if the hTERT mRNA in the sample that comprises the β sequence comprises at least 59% of hTERT mRNA in the sample.