Evaluation of microsatellite instability in women with epithelial ovarian cancer

Abstract

The function of microsatellite instability (MSI) and the optimal panel of markers for epithelial ovarian cancer (EOC) are not well established. This study aimed to use the National Cancer Institute (NCI) markers BAT25, BAT26, D2S123, D5S346 and D17S250 to evaluate MSI in patients with ovarian serous cystadenocarcinoma, compared with ovarian serous cystadenoma and normal ovaries. A total of 37 patients were divided into three groups, as follows: cystadenocarcinoma (n=13), cystadenoma (n=10) and normal ovaries (n=14). DNA was extracted with TRIzol and quantified by spectrophotometry. MSI was evaluated by polymerase chain reaction (PCR), and classified as high (MSI-H), low (MSI-L) or stable (MSS). FIGO staging was I/II in 23.1% and III/IV in 76.9% of the cystadenocarcinoma group. Polymorphisms were found using at least one marker in 32 women, and were observed with D2S123 (83.7%), D17S250 (81.1%), D5S346 (72.9%), BAT25 (21.6%) and BAT26 (16.2%) markers. In the cystadenocarcinoma group, BAT25, BAT26, D2S123, D5S346 and D17S250 markers were positive in 30.8, 76.9, 53.8, 69.2 and 69.2% of patients, respectively. The same markers were positive in 30, 50, 40, 60 and 30% of the cystadenoma group, and 50, 71.4, 71.4, 64.3 and 63.3% in the normal ovary group, respectively. MSI-H was present in 84.6, 60 and 78.6% of the cystadenocarcinoma, cystadenoma and normal patients, respectively. MSI-L was detected in 0, 30 and 7.1%, and MSS was identified in 15.4, 10 and 14.3% of the cystadenocarcinoma, cystadenoma and normal patients, respectively. The frequency of MSI in both benign epithelial ovarian neoplasms and in normal ovaries was high, as well as in EOC, with no statistically significant difference between the groups. This suggests that MSI may arise as a consequence of the ovulatory process, and not solely as a feature of malignant ovarian tumors.

Figure 1

Polyacrylamide gel electrophoresis (7.5%) of NCI markers (A, C, E) and peripheral blood samples (B, D, F) of patients. (A, B) Cystadenocarcinoma, (C, D) cystadenoma, and (E, F) normal ovarian tissue. Columns 1 and 6, BAT25; 2 and 7, BAT26; 3 and 8, D2S123; 4 and 9, D5S346; and 5 and 10, D17S250. BAT25, BAT26, D2S123 (A), D2S123 (C) and BAT26, D2S123, D5S346, D17S250 (E) polymorphic alleles are present in the ovarian tissues and absent in the peripheral blood samples (B, D, F), characterized as MSI-H, MSI-L and MSI-H, respectively. L, 100-bp DNA ladder.

Figure 2

NCI marker frequency in cystadenocarcinoma (CystadenoCa), cystadenoma (Cyst) and normal ovaries.

Figure 3

Frequency of microsatellite instability (MSI). High (MSI-H), low (MSI-L) or stable (MSS).

Figure 4

qPCR for MLH1 and MSH2 genes in the normal ovary, cystadenoma (Cyst) and cystadenocarcinoma (EOC) groups. There was no difference in gene expression levels between groups, when compared by the Kruskal-Wallis test.