Agnieszka Timorek

PIK3CA amplification associates with resistance to chemotherapy in ovarian cancer patients.

PI3K/AKT signalling pathway controls important cellular processes such as the cell proliferation and apoptosis. PIK3CA gene encoding a catalytic subunit of the PI3K is mutated and/or amplified in various neoplasms, including ovarian cancer. We aimed to evaluate PIK3CA alterations and their clinical importance in ovarian cancer patients. Molecular analysis was performed on 117 ovarian carcinomas with the use of qPCR, SSCP and sequencing. In a group of 98 patients with complete clinical data, 62 patients were treated with standard taxane-platinum regimens and 36 patients with platinum-cyclophosphamide regimens. A multivariate analysis was performed by the Cox’s and logistic regression models. PIK3CA mutations occurred in 5/117 (4.3%) carcinomas, exclusively in the endometrioid and clear cell types (P=0.0002); they were also associated with low FIGO stage (P=0.0003), low tumor grade (P=0.045) and early patient’s age at diagnosis (P=0.0005). The PIK3CA amplification (predominantly a low-level) was found in 28/117 (24%) ovarian carcinomas. It was more frequent in TP53 mutant tumors (P=0.012) and tended to associate with high pAKT expression (P=0.061). The PIK3CA amplification strongly diminished odds of complete remission (OR=0.25, P=0.033) and platinum sensitive response (PS, OR=0.12, P=0.004) in the taxane-platinum treated patients. The odds of PS were also much lower in all patients with the PIK3CA amplification evaluated together, regardless of the treatment applied (OR=0.18, P=0.001). Our results suggest that PIK3CA amplification may be a marker predicting ovarian cancer response to chemotherapy.

Androgen, progesterone, and FSH receptor polymorphisms in ovarian cancer risk and outcome

Genes encoding hormone receptors are among candidate genes modulating the risk of ovarian cancer. We aimed to assess a frequency of PGRG+331A, FSHRAla307Thr, and FSHRSer680Asn polymorphic variants, and the length of (CAG)n and (GGN)n repeat tracts in the androgen receptor gene (AR) with respect to ovarian cancer risk and outcome. We genotyped 215 ovarian cancer patients and 352 unaffected control subjects. Statistical analysis was performed with the logistic regression model with adjustment for age. Clinical importance of the polymorphic variants was evaluated in multivariate models on 69 patients treated with taxane-platinum chemotherapy, with respect to TP53 status. Longer AR (GGN)n and (CAG)n repeat tracts decreased the risk of ovarian cancer. For (GGN)n, each additional repeat decreased the risk by 17% (P=0.011) or 27% (P=0.002), while the presence of at least 23 repeats decreased the risk by 41% (P=0.032) or 68% (P=0.008), for the shorter or longer allele respectively. The risk of disease was also decreased by 11% with each additional (CAG)n repeat (P=0.006 for the longer allele). FSHRAla307Ala or FSHRSer680Ser polymorphisms increased ovarian cancer risk by 1.8 times (P=0.042). In all 69 patients, longer AR (CAG)n repeats decreased the risk of recurrence (P=0.031). In the group with TP53 accumulation, longer AR (CAG)n repeats decreased the risk of recurrence (P=0.003) and death (P=0.03), while the FSHRSer680Ser polymorphism increased the risk of recurrence (P=0.037). Progesterone receptor polymorphisms analyzed did not show any associations. Our results support both the androgen and gonadotropin hypotheses of ovarian cancer development.

TP53 status and taxane-platinum versus platinum-based therapy in ovarian cancer patients: A non-randomized retrospective study

BackgroundTaxane-platinum therapy (TP) has replaced platinum-based therapy (PC or PAC, DNA damaging chemotherapy) in the postoperative treatment of ovarian cancer patients; however, it is not always effective. TP53 protein plays a differential role in response to DNA-damaging agents and taxanes. We sought to define profiles of patients who benefit the most from TP and also of those who can be treated with PC.MethodsWe compared the effectiveness of PC/PAC (n = 253) and TP (n = 199) with respect to tumor TP53 accumulation in ovarian cancer patients with FIGO stage IIB-IV disease; this was a non-randomized retrospective study. Immunohistochemical analysis was performed on 452 archival tumors; univariate and multivariate analysis by the Coxs and logistic regression models was performed in all patients and in subgroups with [TP53(+)] and without TP53 accumulation [TP53(-)].ResultsThe advantage of taxane-platinum therapy over platinum-based therapy was seen in the TP53(+), and not in the TP53(-) group. In the TP53(+) group taxane-platinum therapy enhanced the probability of complete remission (p = .018), platinum sensitivity (p = .014), platinum highly sensitive response (p = .038) and longer survival (OS, p = .008). Poor tumor differentiation diminished the advantage from taxane-platinum therapy in the TP53(+) group. In the TP53(-) group PC/PAC was at least equally efficient as taxane-platinum therapy and it enhanced the chance of platinum highly sensitive response (p = .010). However, in the TP53(-) group taxane-platinum therapy possibly diminished the risk of death in patients over 53 yrs (p = .077). Among factors that positively interacted with taxane-platinum therapy in some analyses were endometrioid and clear cell type, FIGO III stage, bulky residual tumor, more advanced age of patient and moderate tumor differentiation.ConclusionOur results suggest that taxane-platinum therapy is particularly justified in patients with TP53(+) tumors or older than 53 years. In the group of patients ≤53 yrs and with TP53(-) tumors platinum-based therapy is possibly equally efficient. We provide hints for planning randomized trials to verify these observations.

TP53, BCL-2 and BAX analysis in 199 ovarian cancer patients treated with taxane-platinum regimens.

OBJECTIVE In cell line studies, BCL-2 and BAX proteins interfere with cancer response to taxanes. This issue has not received much attention with regard to taxane-platinum (TP)-treated ovarian cancer patients. METHODS We evaluated prognostic/predictive significance of BCL-2 and BAX with regard to TP53 status. Immunohistochemical analysis was performed on 199 ovarian carcinomas FIGO stage IIB-IV treated with TP; the results were analyzed by the Cox and logistic regression models. RESULTS Clinicopathological parameters (residual tumor size, FIGO stage and/or tumor grade, but not patients age) were the only or the strongest predictors of patients outcome. Platinum highly sensitive response showed a positive association with TP53 accumulation (p=0.045). As in our previously published analysis on platinum-cyclophosphamide-treated group, complete remission showed a borderline negative (paradoxic) association with high BAX expression in the whole group (p=0.058) and with BCL-2 expression in the TP53(-) group (p=0.058). CONCLUSION Our results suggest that TP53, BCL-2 and BAX proteins carry some predictive potential in taxane-platinum-treated ovarian cancer patients, auxiliary to clinicopathological factors. We have confirmed on another patient group that clinical importance of BCL-2 may depend on TP53 status.

The significance of c.690G>T polymorphism (rs34529039) and expression of the CEBPA gene in ovarian cancer outcome

The CEBPA gene is known to be mutated or abnormally expressed in several cancers. This is the first study assessing the clinical impact of CEBPA gene status and expression on the ovarian cancer outcome. The CEBPA gene sequence was analyzed in 118 ovarian cancer patients (44 platinum/cyclophosphamide (PC)-treated and 74 taxane/platinum (TP)-treated), both in tumors and blood samples, and in blood from 236 healthy women, using PCR-Sanger sequencing and Real-Time quantitative PCR (qPCR)-based genotyping methods, respectively. The CEBPA mRNA level was examined with Reverse Transcription quantitative PCR (RT-qPCR). The results were correlated to different clinicopathological parameters. Thirty of 118 (25.4%) tumors harbored the CEBPA synonymous c.690G>T polymorphism (rs34529039), that we showed to be related to up-regulation of CEBPA mRNA levels (p=0.0059). The presence of the polymorphism was significantly associated with poor prognosis (p=0.005) and poor response to the PC chemotherapy regimen (p=0.024). In accordance, elevated CEBPA mRNA levels negatively affected patient survival (p<0.001) and tumor response to the PC therapy (p=0.014). The rs34529039 SNP did not affect the risk of developing ovarian cancer. This is the first study providing evidence that the c.690G>T, p.(Thr230Thr) (rs34529039) polymorphism of the CEBPA gene, together with up-regulation of its mRNA expression, are negative factors worsening ovarian cancer outcome. Their adverse clinical effect depends on a therapeutic regimen used, which might make them potential prognostic and predictive biomarkers for response to DNA-damaging chemotherapy.

Prognosis of patients with BRCA1-associated ovarian carcinomas depends on TP53 accumulation status in tumor cells

OBJECTIVE TP53 mutation is the most frequent molecular event in BRCA1-associated ovarian carcinomas. TP53 status may be a confounding factor in the evaluation of clinical importance of other proteins. We aimed to evaluate the clinical significance of BRCA1 mutations with respect to the TP53 accumulation status in 159 high-grade ovarian carcinomas. METHODS Statistical analyses were done with the Kaplan-Meier method, log-rank test, the Coxs and logistic regression models for all patients, and in subgroups with and without TP53 accumulation (TP53+ and TP53-, respectively). RESULTS Forty of 159 ovarian carcinomas (25.2%) were diagnosed in patients with BRCA1 germline mutations; 102 tumors (64.2%) were TP53+ and 57 (37.8%) were TP53-. Among patients with TP53+ carcinomas, BRCA1 carriers had increased odds of recurrence compared with sporadic cases (HR 2.25, P=0.003; median disease-free survival time 7.7 vs. 18.4months, respectively). In the smaller TP53- subgroup, BRCA1 mutation reduced the risk of death by 46% (HR 0.54, P=0.099, median overall survival time 42.7 vs. 28.1months), but beyond the border of significance. When the TP53 status was not taken into account, BRCA1 mutations did not show any significance, however, there was a trend toward increased odds of complete remission for women with BRCA1 mutations compared to non-carriers (OR 2.47, P=0.064). Taxane-platinum therapy showed advantage over the platinum-cyclophosphamide one in the entire group of patients and in the TP53+ subgroup. CONCLUSIONS Our results suggest that the TP53 accumulation status determines the prognosis of BRCA1 mutation carriers with high-grade ovarian carcinomas.

Using our own developed stent in the palliative treatment of obstruction in the left half of the colon due to ovarian cancer

OBJECTIVES An assessment of implantation efficacy and safety of self-developed self-expanding stent in patients with an ovarian cancer induced by intestinal obstruction. MATERIAL AND METHODS The study of the stenting efficacy and safety was realized prospectively. The group consisted of 13 patients with left half colon obstruction due to an inoperable metastatic ovarian carcinoma. All the patients had a histopathologically diagnosed ovarian carcinoma and were treated in the past both surgically and systemically. Stenting was preceded by a Computed Tomography (CT) scan confirming and locating the obstruction. Patients with a multilevel intestinal obstruction were disqualified. RESULTS Nine stents were implanted in the rectosigmoid; 4 stents were implanted in an externally compressed rectum. One migration of implanted stent was observed. In one case 2 stents were implanted due to an insufficient coverage of the stricture. The decompression of the obstruction of the gastrointestinal tract was achieved in 11 patients (85%). CONCLUSIONS 1) The implantation of our own developed, self-expanding stent is effective and safe. 2) The implantation of the stent in patients with an inoperable ovarian cancer causing an obstruction of the gastrointestinal tract is an effective procedure limiting postoperative complications and improving life comfort by avoiding stoma.

The role of mathematical models and scoring systems in detecting dysgerminoma in a young patient

A 20-year-old Caucasian woman presented at the emergency unit with a complaint of lower abdominal pain, which was had been present for a few months but worsened on that day. The patient had no family history of cancer. She denied weight loss, vomiting, diarrhea, and constipation. The patient is a virgin, with irregular and heavy menses (6–7 days, every 33–45 days). Transabdominal ultrasound was performed by an inexperienced examiner using Philips iu 22z apparatus. Gray-scale ultrasound examination revealed an irregular heteroechogenic solid mass (114 × 77 × 72 mm in size) in the pelvis, protruding into the abdomen on the left side (Fig. 1). There were no acoustic shadows and the patient reported no pain during the examination. There was only a small amount of fluid below the mass (Fig. 2). Color Doppler examination of the mass revealed moderate vascularization, assessed as score 3 according to the terms and definitions established by the International Ovarian Tumor Analysis (IOTA) group (Fig. 3). The second ovary was irregular (40 × 28 mm), in close proximity to the pelvic mass with hypoechogenic areas, suggestive of metastases. The examination revealed a hypoechogenic area of 15.5 × 17 mm in the pancreas, suggestive of metastases (Fig. 4). The liver and the kidneys had no focal lesions. The mass was considered as malignant according to the subjective assessment by the inexperienced ultrasound examiner. Ten selected mathematical models and scoring systems, based on clinical features, ultrasound examination and serum concentration of tumor markers, were used to assess the mass by the inexperienced examiner. The risk of Malignancy Algorithm (ROMA) revealed a high (10.24%) risk for ovarian cancer malignancy. Malignant criterion as irregular solid mass in the absence of other benign criteria suggested a malignant nature of the mass, according to the simple rules of the IOTA group. The results of the IOTA logistic regression models LR1 and LR2 were 43.7% and 23.5% respectively, indicating a malignant character of the lesion. The pelvic mass was considered to be malignant according to the risk of malignancy indices RMI I–IV, whose results were 512, 683, 512 and 1366, respectively. The Gynecologic Imaging Report and Data System classified the mass as grade 4, i.e. “probably malignant”. The ADNEX model revealed a 68.2% risk of malignancy, with 27.4% risk of being stage II–IV ovarian cancer. After obtaining the patient’s consent, fertility-sparing surgical operation was performed. The final diagnosis was dysgerminoma, stage IV according to the International Federation of Gynecology and Obstetrics (FIGO) criteria. In the case of our patient, advanced stage of the ovarian cancer might have facilitated the diagnosis as the changes were evident. On the other hand, the rarity of the disease, young age of the patient, and non-specific symptoms may cause diagnostic difficulties. Systemic approach, with the use of clinical, ultrasound and tumor markers, may help not to overlook ovarian cancer even in younger patients when assessed by inexperienced examiners. Figure 1. Irregular solid tumor of the left ovary