Cyclin D3 expression in primary Ta/T1 bladder cancer

Abstract

Cyclin D3 deregulation has recently been reported in bladder cancer but its prognostic significance remains uncertain. A cohort of 159 patients with stage Ta or T1 primary bladder tumours was investigated to determine the significance of cyclin D3 expression in association with other G1-S phase regulators of the cell cycle (p53, p21Waf1, p27kip1, cyclin D1), including tumour proliferation (ki67-MIB1); its association with conventional clinicopathological parameters; and the relationship between cyclin D3 and loss of heterozygosity (LOH) at the 9p21 (p16INK4a locus) chromosome region. The end point of the study was progression-free survival. Cyclin D3, other G1-S phase regulators, and tumour proliferation were investigated by immunohistochemistry and measured by the grid-counting method. To validate the immunohistochemical expression, cyclin D3 was additionally assessed by western blotting in selected cases. LOH at the 9p21 chromosome region (marker D9S171) was assessed in 125 cases using an AB Prism 310 genetic analyser and a set of microsatellite fluorescence-labelled primers. Cyclin D3 overexpression was related to larger tumour size (>5 cm; p < 0.0001) and high tumour proliferation (>10%; p = 0.025). Mean cyclin D3 expression increased with 2004 WHO grading categories in stage Ta (p = 0.035, ANOVA) and stage T1 (p = 0.047, t test) tumours. Cyclin D3 was not related to other clinicopathological parameters, G1-S phase modulators, or 9p21 LOH. Cox's multivariate analysis selected cyclin D3 as an independent predictor of progression-free survival (p = 0.0012, relative risk (RR) = 5.2366) together with tumour size (p = 0.0115, RR = 4.4442) and cyclin D1 (p = 0.0065, RR = 3.3023). Cyclin D3 expression had the highest risk ratio. Our results suggest that expression of cyclin D3 is relevant to the progression-free survival of patients with Ta/T1 bladder carcinomas.

Cyclin D3 gene amplification in bladder carcinoma in situ

“…We found up-regulation of Cyclin D3 gene to be present in about one third of secondary CIS cases. This result agrees with our previous work showing Cyclin D3 protein upregulation in high-grade bladder carcinoma [24,25,27]. In addition, we have observed that none of primary CIS cases entering the current study had Cyclin D3 gene amplification, raising the question if the known differences in aggressiveness between primary and secondary CIS cases could be explained by differences in genetic alterations between these two forms of bladder CIS; this could be related to a higher cell-cycle regulatory genes alteration in secondary CIS.…”

“…Our finding of a strong association of Cyclin D3 gene amplification with recurrence-free and mainly with progression-free survival could indicate that there is an important deregulation of Cyclin D3 gene and protein over-expression in high-grade bladder cancer [24,25,27]. Moreover, it seems that Cyclin D3 alteration is a potentially relevant finding since the use of cell-cycle regulatory proteins as therapeutic targets of their modulators, represents an active field of research [24]. Available modulators of Cyclin D3 expression such Interferon-α and flavopiridol, that can induce cell-cycle arrest by directly inhibiting cyclin dependent kinases and by downregulation of Cyclin D3, might represent a future alternative to other conventional therapeutic approaches in a subset of bladder CIS patients at higher risk of aggressive behavior.…”

“…Our finding of a strong association of Cyclin D3 gene amplification with recurrence-free and mainly with progression-free survival could indicate that there is an important deregulation of Cyclin D3 gene and protein over-expression in high-grade bladder cancer [24,25,27]. Moreover, it seems that Cyclin D3 alteration is a potentially relevant finding since the use of cell-cycle regulatory proteins as therapeutic targets of their modulators, represents an active field of research [24].…”

“…Cyclin D3 gene may be deregulated in a number of human malignant tumors [24][25][26][27][28]. Recent reports suggest that Cyclin D3 gene is deregulated in bladder urothelial carcinoma with no reports concerning CIS [24,25,27]. The status of Cyclin D3 gene in bladder CIS remains unknown.…”

“…The value of other markers for progression in CIS patients, like oncogenes, cell-cycle regulator genes, proliferation, neovascularization markers and growth factors remains to be determined [15][16][17][18][19] Cyclin D3 plays a pivotal role in controlling physiological progression from the G1 to the S phase of the cell cycle. Cyclin D3 gene may be deregulated in a number of human malignant tumors [24][25][26][27][28]. Recent reports suggest that Cyclin D3 gene is deregulated in bladder urothelial carcinoma with no reports concerning CIS [24,25,27].…”