ESR1 and Cervical Cancer

• US 20090023137A1
• Filed: 07/07/2005
• Published: 01/22/2009
• Est. Priority Date: 07/16/2004
• Status: Abandoned Application

• Current Assignee: Oncomethylome Sciences SA
• Original Assignee: Oncomethylome Sciences SA
• Inventors: Van Der Zee, Ate, Wisman, Bea
• Application Number:
  US11/632,493
• Publication Number:
  US 20090023137A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/154 Methylation markers
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays

Abstract

The present invention provides methods and kits for detecting susceptibility to, or the incidence of, cervical cancer in a sample comprising determining the methylation status of the ESR1 gene, optionally as part of a panel of genes. Also provided are methods and kits which involve determining the expression levels of genes including the ESR1 gene in order to diagnose cervical cancer.

Claims

1. A method of detecting susceptibility to, or the incidence of, cervical cancer in a sample comprising determining the methylation status of ESR1.

2. The method according to claim 1, comprising determining whether the ESR1 gene is hypermethylated.

3. The method according to claim 2, wherein ESR1 forms one of a panel of genes whose methylation status is linked to the incidence of cervical cancer.

4. The method according to claim 3, wherein the panel of genes also comprises at least one of the following genes:

DAP-kinase, APC, TIMP-3, RAR-beta, CALCA, TSLC1, TIMP-2, DcR1, DcR2, BRCA1, p15, Rassf1A, MLH1, MGMT.

5. The method according to claim 4, wherein the panel of genes also comprises DAP-kinase.

6. The method according to claim 1, wherein the method comprises determining the methylation status of a total of three genes.

7. The method according to claim 6, wherein the three genes comprise ESR1 and two genes selected from DAPK, APC, TIMP-3, CALCA and RAR-beta.

8. The method according to claim 1, wherein the method comprises determining the methylation status of a total of four genes.

9. The method according to claim 8, wherein the four genes comprise ESR1 and three genes selected from DAPK, APC, TIMP 3, CALCA and RAR-beta.

10. The method according to claim 1, wherein the method comprises determining the methylation status of a total of five genes.

11. The method according to claim 10, wherein the five genes comprise ESR1 and four genes selected from DAPK, APC, TIMP-3, CALCA and RAR-beta.

12. The method according to claim 1, wherein the method allows at least 90% detection of cervical cancer.

13. The method according to claim 1, wherein the methylation status of the gene(s) is determined using methylation specific PCR (MSP).

14. The method according to claim 13, wherein the methylation status of ESR1 is determined using primers comprising the sequence of SEQ ID NO:

1 and SEQ ID NO;

2.

15. The method according to claim 13, wherein the methylation status of ESR1 is determined using primers consisting essentially of the sequence of SEQ ID NO:

1 and SEQ ID NO;

2.

16. The method according to claim 1, wherein the methylation status of the gene(s) is determined using quantitative methylation specific PCR (QMSP).

17. The method according to claim 16, wherein the methylation status of ESR1 is determined using primers comprising the nucleotide sequence of SEQ ID NO:

1 and SEQ ID NO;

2 and a probe comprising the sequence of SEQ ID NO;

3.

18. The method according to claim 16, wherein the methylation status of ESR1 is determined using primers consisting essentially of the nucleotide sequence of SEQ ID NO:

1 and SEQ ID NO;

2 and a probe consisting essentially of the sequence of SEQ ID NO;

3.

19. The method according to claim 1, wherein the sample is a cervical scraping.

20. A kit for use in detecting susceptibility to, or the incidence of, cervical cancer in a cervical sample from a patient comprising;

a) gene specific primers for determining the methylation status of the ESR1 gene; and

b) an Ayre'"'"'s spatula and/or an endocervical brush for removing cervical cells from the patient.

21. The kit according to claim 20, further comprising a gene specific probe for the ESR1 gene.

22. The kit according to claim 20, wherein the gene specific primers comprise those comprising the sequence of SEQ ID NO:

1 and SEQ ID NO;

2.

23. The kit according to claim 21, wherein the gene specific probe comprises that comprising the sequence of SEQ ID NO:

3.

24. The kit according to claim 20, wherein the gene specific primers comprise those consisting essentially of the sequence of SEQ ID NO:

1 and SEQ ID NO;

2.

25. The kit according to claim 21, wherein the gene specific probe comprises that consisting essentially of the sequence of SEQ ID NO:

3.

26. The kit according to claim 20, further comprising DNA isolation reagents.

27. The kit according to claim 20, further comprising enzymes for amplification of DNA.

28. The kit according to claim 20, further comprising sodium bisulphite.

29. The kit according to claim 20, further comprising suitable buffers.