• Mantle cell lymphoma (MCL) is a subtype of non-Hodgkin’s lymphoma characterized by poor prognosis and a median survival of approximately 3 years. 
• On the basis of morphology and immunophenotype alone, mantle cell lymphoma is difficult to distinguish from indolent lymphomas and leukemias. 
• Cytogenetically, a t(11:14)(q13;q32) is associated with 75% of mantle cell lymphomas. 
• PCR for the major translocation cluster (MTC) region of the bcl-1 is positive in only 30-40% of cases. 
• Southern blot analysis detects translocations in approximately 50% to 70% of the cases. 
• Translocation breakpoints are scattered within the approximately 120-kb bcl-1 region adjacent to cyclin D1, whereas part of them are clustered in the MTC. 
• The translocation leads to overexpression of cyclin D1 due to juxtaposition of the Ig heavy chain gene enhancer on 14q32 to the cyclin D1 gene on 11q13. 

• Cyclin D1 amplification is common in many types of cancer, second only to the HER2/neu (erb-b2) oncogene. 
• The LSI Cyclin D1 is approximately 300 kb in size. 
• This probe may be used to determine the copy number of the Cyclin D1 locus, or as an enumerator probe for chromosome 11 in interphase and metaphase cells. 
• 200 cells will be counted. The result will be expressed as the ratio of Cyclin D1/CEP11. 

Print Date: 06/04/1999                Name: ##############
Location: DICT                         Hospital Number:  (00000)#########
Attending : #############         Age:        50 YRS  Sex: F

CYTOPATHOLOGY  -  PHONE:   (713)794-5625
LYMPHOPROLIFERATIVE DISORDER FISH ANALYSIS

ACCESSION # : FI-99-0060 
COLLECTION DATE: 5/27/1999

RESULTS:

SAMPLE TYPE: Bone marrow 
                                           Signals
Probe                        1         2         3        4        5          6       7         8       >8
11CEP(%)                 1        98       1        0        0          0       0         0        0 
Cyclin D1(%)           0       13.5     87      0        0          0       0         0        0
200 cells counted

For Cyclin D1 test:
Ratio  = Total Cyclin D1/Total 11CEP =  287/200 = 1.43
Normal range: Mean ratio +/- SD = 0.99 +/- 0.09.   95% Confident Interval: (0.96, 1.02)
Ratio for Mantle cell positive control: 1.49
Ratio for Mantle cell negative control: 1.00

ISCN DIAGNOSIS: nuc ish 11cen(D11Z1 x 2), 11q13(CCND1 x 3) [200]

INTERPRETATION:

Elevated ratio Cyclin D1: total 11 CEP at 1.43 with 87% cells showing evidence of Cyclin D1 translocation consistent with extensive involvement by mantle cell lymphoma in bone marrow.

Notes: Based on mixing experiments, in peripheral blood, bone marrow, or lymph-node specimens, the level of detectable translocation t(11;14)(q13;q32) cells is considered positive if 5% or higher numbers of translocated cells are presented. 

The in situ hybridization (ish) technique was performed using a Vysis LSI Cyclin D1/CEP11 probe. A total of 200 interphases were analyzed. 

In situ hybridization is for investigational use only and should not be used for diagnostic purposes without confirmation by another proven procedure. This test does not rule out abnormalities other than the one for which the probe was designed.

REFERENCE:  R.L.Katz, J.Gu, L.Pasco-Miller, H.Z.Zhang, N.Caraway, and L.J.Medeiros. Detection of Cyclin D1 Translocation by Fluorescence in-situ Hybridization (FISH) is Highly Specific and Sensitive in the Diagnosis of Mantle Cell Lymphoma (MCL) Modern Pathology Vol.12.1999. 

Date:    Verified by:
CAC     (Electronic Signature)

           * Result will be reported on the Cerner in about a week after sample received.
 
 

Figure 1. Interphase FISH from a normal cell line. Cells showing 2 green signals (Centromere 11) and 2 orange signals (Cyclin D1 at chromosome 11q13).	Figure 2. Interphase FISH from a non-MCL patient. Cells showing 2 green signals (Centromere 11) and 2 orange signals (Cyclin D1 at chromosome 11q13).
Figure 3. Another interphase FISH from a non-MCL patient. Cells showing 2 green signals (Centromere 11) and 2 orange signals (Cyclin D1 at chromosome 11q13).	Figure 4. Interphase FISH from a cell line with cytogenetically detectable t(11;14)(q13;q32). Most of cells showing 2 green signals (Centromere 11) and 3 orange signals (Cyclin D1 at chromosome 11q13).
Figure 5. Interphase FISH from a MCL patient with tetraploid cell population. Most of the cells showing 4 green signals (Centromere 11) and 5  orange signals (Cyclin D1 at chromosome 11q13).	Figure 6. Interphase FISH from a blastoid MCL patient. Cells are showing 2 green signals (Centromere 11) and 3 or more orange signals (Cyclin D1 at chromosome 11q13).