Methods of cytodiagnostic staging of neoplasia and squamous cell carcinoma

1. A method for diagnosing whether an epithelial tissue is an abnormal tissue comprising

determining an expression pattern for PML in the epithelial tissue;

determining an expression pattern for nuclear bodies in the epithelial tissue;

determining a loss of SUMO-1 colocalization with nuclear bodies comprising PML in the epithelial tissue; and

comparing the expression pattern for PML, the expression pattern for nuclear bodies, and the loss of SUMO-1 colocalization with nuclear bodies comprising PML with that of a normal tissue, where loss of SUMO-1 colocalization with nuclear bodies comprising PML indicates abnormal tissue.

2. The method of claim 1 , wherein the expression patterns of the normal tissue is determined from a normal tissue sample.

3. The method of claim 1 , wherein the normal tissue exhibits

(a) a strong expression pattern for PML in the basal layer of the epithelial tissue;

(b) basal cells having a consistent PML pattern with an average of about 5 round nuclear bodies per nucleus and an area of about 0.25 μm 2 ;

(c) a weak expression pattern for PML in the suprabasal and intermediate layers of the epithelial tissue as compared to (a);

(d) no expression of PML in the superficial layer of the epithelial tissue;

(e) PML expression in the stroma of the epithelial tissue;

(f) a PML expression pattern in the vascular cells of the epithelial tissue similar to (a);

(g) about all of the nuclear bodies comprising PML colocalize with SUMO-1; or

(h) a combination thereof.

4. The method of claim 1 , wherein the abnormal tissue exhibits

(a) PML expression restricted to the lower half of the epithelium of the epithelial tissue;

(b) nuclear bodies having irregular shapes;

(c) about 9 to about 10 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(d) nuclear bodies having an area of about 0.52 μm 2 to about 0.54 μm 2 ;

(e) about 95% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof.

5. The method of claim 1 , wherein the abnormal tissue is cervical intraepithelial neoplasia.

6. The method of claim 4 , wherein the abnormal tissue is mild dysplasia, moderate dysplasia, or a combination thereof.

7. The method of claim 1 , wherein the epithelial tissue exhibits

(a) PML expression throughout the epithelial tissue;

(b) nuclear bodies having irregular shapes and nuclear bodies having track-like structures;

(c) about 9 to about 10 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(d) nuclear bodies having an area of about 0.55 μm 2 ;

(e) about 93% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof which is abnormal tissue.

8. The method of claim 7 , wherein the abnormal tissue is Type A severe dysplasia.

9. The method of claim 1 , wherein the epithelial tissue exhibits

(a) downregulated PML expression;

(b) about 5 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(c) nuclear bodies having an area of about 0.33 μm 2 ;

(d) about 49% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(e) a combination thereof, which is abnormal tissue.

10. The method of claim 9 , wherein the abnormal tissue is Type B severe dysplasia.

11. The method of claim 1 , wherein the epithelial tissue exhibits

(a) upregulated PML expression;

(b) variation in the number and size of nuclear bodies;

(c) about 9 nuclear bodies in the epithelial tissue;

(d) nuclear bodies having an area of about 0.73 μm 2 ;

(e) about 18% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof which is abnormal tissue.

12. The method of claim 11 , wherein the abnormal tissue is well-differentiated cervical squamous cell carcinoma.

13. The method of claim 1 , wherein the abnormal tissue exhibits

(a) downregulated PML expression;

(b) about 5 nuclear bodies in the epithelial tissue;

(c) nuclear bodies having an area of about 0.37 μm 2 ;

(d) about 10% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(e) a combination thereof.

14. The method of claim 13 , wherein the abnormal tissue is poorly-differentiated cervical squamous cell carcinoma.

15. The method of claim 1 , wherein SUMO-1 colocalization with about 95% of nuclear bodies comprising PML indicates that the abnormal tissue is mild dysplasia, moderate dysplasia, or a combination thereof.

16. The method of claim 1 , wherein SUMO-1 colocalization with about 93% of nuclear bodies comprising PML indicates that the abnormal tissue is Type A severe dysplasia.

17. The method of claim 1 , wherein SUMO-1 colocalization with about 49% of nuclear bodies comprising PML indicates that the epithelial tissue is abnormal tissue which is Type B severe dysplasia.

18. The method of claim 1 , wherein SUMO-1 colocalization of about 18% indicates that the epithelial tissue is abnormal tissue which is well-differentiated cervical squamous cell carcinoma.

19. The method of claim 1 , wherein SUMO-1 colocalization of about 10% indicates that the epithelial tissue is abnormal tissue which is poorly-differentiated cervical squamous cell carcinoma.

20. The method of claim 1 , wherein the PML expression pattern and SUMO-1 colocalization nuclear bodies comprising PML are fluorescently detected in a paraffin embedded tissue biopsy of the epithelial tissue.

21. A method for diagnosing whether a subject has mild dysplasia, moderate dysplasia, Type A severe dysplasia, Type B severe dysplasia, well differentiated cervical squamous cell carcinoma, or poorly-differentiated cervical squamous cell carcinoma comprising

determining an expression pattern for PML an epithelial tissue sample from the subject;

determining an expression pattern for nuclear bodies in the epithelial tissue sample;

determining a loss of SUMO-1 colocalization with nuclear bodies comprising PML in the epithelial tissue sample; and

determining whether the expression pattern for PML, the expression pattern for nuclear bodies, and the loss of SUMO-1 colocalization with nuclear bodies comprising PML of the epithelial tissue sample is consistent with expression patterns of mild dysplasia, moderate dysplasia, Type A severe dysplasia, Type B severe dysplasia, cervical squamous cell carcinoma, or poorly-differentiated cervical squamous cell carcinoma.

22. The method of claim 21 , further comprising determining SUMO-1 colocalization with nuclear bodies comprising PML in the tissue sample, wherein

SUMO-1 colocalization with about 95% of nuclear bodies comprising PML indicates that the subject has mild dysplasia, moderate dysplasia, or a combination thereof;

SUMO-1 colocalization with about 93% of nuclear bodies comprising PML indicates that the subject has Type A severe dysplasia;

SUMO-1 colocalization with about 49% of nuclear bodies comprising PML indicates that the subject has Type B severe dysplasia;

SUMO-1 colocalization with about 18% of nuclear bodies comprising PML indicates that the subject has cervical squamous cell carcinoma; and

SUMO-1 colocalization with about 10% of nuclear bodies comprising PML indicates that the subject has poorly-differentiated cervical squamous cell carcinoma.

23. The method of claim 21 , wherein

(a) PML expression restricted to the lower half of the epithelium of the epithelial tissue;

(b) nuclear bodies having irregular shapes;

(c) about 9 to about 10 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(d) nuclear bodies having an area of about 0.52 μm 2 to about 0.54 μm 2 ;

(e) about 95% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof indicates that the subject has mild dysplasia, moderate dysplasia, or a combination thereof.

24. The method of claim 21 , wherein

(a) PML expression throughout the epithelial tissue;

(b) nuclear bodies which have inconsistent shapes and track like structures;

(c) about 9 to about 10 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(d) nuclear bodies having an area of about 0.55 μm 2 ;

(e) about 93% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof indicates that the subject has Type A severe dysplasia.

25. The method of claim 21 , wherein

(a) downregulated PML expression;

(b) about 5 nuclear bodies in the basal layer and upper layer of the epithelial tissue;

(c) nuclear bodies having an area of about 0.33 μm 2 ;

(d) about 49% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(e) a combination thereof indicates that the subject has Type B severe dysplasia.

26. The method of claim 21 , wherein

(a) upregulated PML expression;

(b) variation in the number and size of nuclear bodies;

(c) about 9 nuclear bodies in the epithelial tissue;

(d) nuclear bodies having an area of about 0.73 μm 2 ;

(e) about 18% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(f) a combination thereof indicates that the subject has well-differentiated cervical squamous cell carcinoma.

27. The method of claim 21 , wherein

(a) downregulated PML expression;

(b) about 5 nuclear bodies in the epithelial tissue; (c) nuclear bodies having an area of about 0.37 μm 2 ;

(d) about 10% of the nuclear bodies comprising PML colocalize with SUMO-1; or

(e) a combination thereof indicates that the subject has poorly-differentiated cervical squamous cell carcinoma.

28. The method of claim 21 , wherein the subject is mammalian.

29. The method of claim 21 , wherein the subject is human.

30. The method of claim 1 , wherein the epithelial tissue is cervical epithelial tissue.

31. A method for diagnosing whether a cervical epithelial tissue is an abnormal tissue comprising

determining an expression pattern for PML in the cervical epithelial tissue;

determining an expression pattern for nuclear bodies in the cervical epithelial tissue;

determining a loss of SUMO-1 colocalization with nuclear bodies comprising PML in the cervical epithelial tissue; and

comparing the expression pattern for PML, the expression pattern for nuclear bodies, and the loss of SUMO-1 colocalization with nuclear bodies comprising PML with that of a normal tissue, where loss of SUMO-1 colocalization with nuclear bodies comprising PML indicates abnormal tissue.