Expression of stem cell marker CD44 in prostate cancer biopsies predicts cancer grade in radical prostatectomy specimens.

Cancer stem cells play an important role in development and progression of many cancer types including prostate adenocarcinoma. We used a stem cell marker CD44 to evaluate the prevalence of prostate cancer stem cells in prostate biopsies and in matched radical prostatectomy specimens. We tested both types of specimen for the existence of a correlation between the immunohistochemical expression of CD44 and Gleason grade, pathological stage (pT) according to TNM, patient age and preoperative plasma PSA levels in 52 patients. We found a positive correlation between the expression of CD44 in cancer cells from prostate biopsies and in matched radical prostatectomy specimens. We also observed that higher level of CD44 expression in cancer cells correlated with lower Gleason score, both in prostate biopsies and in radical prostatectomies. To the best of our knowledge we showed for the first time, that the level of CD44 expression in prostate biopsies correlates with that observed in matched radical prostatectomy specimens. Since the level of CD44 expression was shown to predict a response to anti cancer therapy in several types of human tumors, CD44 assessment might support a clinical decision making process in prostate cancer patients.

Two cases of metachronic tumours: cancers of the ovary and endometrium in young women. Case presentation and review.

Ovarian and endometrial cancers seldom develop in females under 40 years of age. Manifestation of metachronic cancers before the age of 40 is of casuistic interest. Two cases are presented in whom metachronic cancers were detected: the first localisation involved the ovary, and the second the endometrium. One of the patients had earlier delivered a baby with multiple hereditary defects and was diagnosed with secondary infertility. The second patient was diagnosed with primary infertility. Immunohistochemical tests disclosed the presence of alpha and beta type estrogen receptors and progesterone receptors in the tumour cells of the ovary and endometrium. Neither of the patients carried mutations in the BRCA 1 or NOD 2 genes.

Evaluation of the sentinel node biopsy in colorectal carcinoma including the results of immunohistochemical examinations.

AIM: To evaluate the method and results of sentinel node biopsy including immunohistochemical examinations in resectable colorectal cancer. MATERIAL AND METHODS: From April 2004 to April 2005, sentinel node biopsy was carried out with the dye method in 27 patients operated on for colorectal cancer. The standard examination of sentinel nodes consisted in the evaluation of individual H&E-dyed specimens from bisection of the node. The negative sentinel nodes were examined with the use of immunohistochemistry against cytokeratins AE1/AE3. FINDINGS: The sentinel node was identified in 25 patients (92.6%). In a routine histopathological examination it included metastases in 3 cases. The sentinel node was clean in one patient whereas other regional nodes resected "en bloc" with the tumour included metastases. The sensitivity of the method was 75%, and the number of falsely negative results was 25%. Metastases (micrometastases) in the sentinel node were found in 2 other patients (8%) in the immunohistochemical examination. This examination did not change the results of the analysis in the patient with positive non-sentinel nodes and with the negative sentinel node in H&E dyeing. Nevertheless, the sensitivity of the method rose to 83.3% and the number of falsely negative results dropped to 16.7%. CONCLUSIONS: 1. The sentinel node biopsy using the dye method is a safe and relatively easy technique showing a high success rate (92.6%). 2. Low sensitivity of the method, as reflected in the literature, may result from lack of extended histopathological examinations performed on the sentinel node (e.g. immunohistochemistry). 3. Further research is necessary to determine the role and importance of the sentinel node biopsy in colorectal cancer.

BRCA1-positive breast cancers in young women from Poland.

We identified 4316 unselected incident cases of early-onset breast cancers (<51 ears of age at diagnosis) in 18 Polish hospitals between 1996 and 2003. We were able to obtain a blood sample for DNA analysis from 3472 of these (80.4%). All cases were tested for the presence of three founder mutations in BRCA1. The proportion of cases with a BRCA1 mutation was 5.7%. The hereditary proportions were higher than this for women with breast cancer diagnosed before age 40 (9%), for women with cancer of medullary or atypical medullary histology (28%), for those with bilateral cancer (29%) or with a family history of breast or ovarian cancer (13%). It is reasonable to offer genetic testing to women with early-onset breast cancer in Poland.

No evidence of HTLV-I infection in patients with mycosis fungoides and Sezary syndrome.

The involvement of human T-cell lymphotropic virus type I (HTLV-I) in the etiology of cutaneous T-cell lymphomas (CTCL) is still controversial. The aim of the study was to evaluate the role of HTLV-I in the pathogenesis of mycosis fungoides (MF) and Sezary syndrome (SS) in Polish patients. The studied group consisted of 42 patients with MF, 5 with SS and 25 with chronic dermatitis. DNA was extracted from snap-frozen and paraffin-embedded skin biopsies and from peripheral blood. Polymerase chain reaction (PCR or nested PCR) was carried out for amplification of different regions of HTLV-I genome. Primer sets flanking pX, p 19, U5, tax and pol genes were used in the investigation. The presence of HTLV-I antibody was examined in 46 sera samples with the use of anti-HTLV-I/II EIA test. HTLV-I antibodies were not detected in any collected sera samples. PCR with two primer sets homologous to the pX region of HTLV-I showed negative results in all samples investigated. To confirm these results two other primer pairs specific for U5 and gag regions were designed. With these primer pairs no PCR product, except that in positive control, was observed. For more sensitive amplification a nested-PCR with pol and tax specific primers was performed. HTLV-I probably does not play an important role in the pathogenesis of MF in Polish patients.

Evaluation of clinical significance of TP53, BCL-2, BAX and MEK1 expression in 229 ovarian carcinomas treated with platinum-based regimen.

In cell line studies, BCL-2, BAX, as well as novel MEK1 protein levels have strong influence on ovarian cancer response to cisplatin-based chemotherapy. However, such associations have not been demonstrated clinically. We evaluated prognostic/predictive significance of these proteins with regard to TP53 status. Immunohistochemical analysis was performed on 229 ovarian carcinomas FIGO stage IIB-IV treated with platinum-based chemotherapy; the results were analysed by the Cox and logistic regression models. Clinical parameters (residual tumour size, patient age, FIGO stage) were the only indicators of overall survival (OS) and the strongest predictors of complete remission (CR). On the other hand, BAX expression was the strongest (P=0.005) or the only (in FIGO IIIC, P=0.02) prognostic indicator of disease-free survival (DFS) in the TP53(+) group. TP53(+) and TP53(-) ovarian carcinomas differed in clinical and molecular prognostic and predictive factors. Another novel finding is that CR was negatively influenced by high BAX expression in all patients group (P=0.047) and by BCL2 expression in the TP53(-) group (P=0.05). High MEK1 expression was associated with endometrioid and clear cell carcinomas (P=0.049); its loss was found with advancing FIGO stage (P=0.002). Our results suggest that binomial TP53 status divides ovarian carcinomas into two biologically distinct groups. BAX expression is an important factor of DFS in the TP53(+) group. BCL-2 and BAX, but not MEK1 expressions have predictive value in ovarian cancer patients treated with platinum-based chemotherapy.

The role of Chlamydia trachomatis infection in cervical cancer development.

Previous Chlamydia trachomatis infection elevates expression of: 1. TGFalpha in CIN I, CIN II, CIN III; 2. HPV 16 in CIN I, CIN II, CIN III and invasive carcinoma; 3. Ki 67 in CIN III and invasive carcinoma. Chronic Chlamydial infection is very often associated with cervical hypertrophy.

[Thyroid cancer in autonomous "hot nodule"]. / Rak tarczycy w autonomicznym "guzku goracym"

17-year-old girl with thyroid tumor of the right lobe is described in this paper. Based on clinical picture and diagnostic criteria the follicular adenoma was suspected pre-operatively as the most likely final diagnosis. Post-operative histopathological evaluation proved the presence of the oxyphilic type of follicular thyroid cancer within the follicular adenoma. The authors analyze the possibilities of the pre-operative discovery of the discussed cancer.

Novel BRCA1 mutations and more frequent intron-20 alteration found among 236 women from Western Poland.

Three different novel BRCA1 mutations, five independent cases of the same 12 bp insertion-duplication in intron-20 and two novel rare BRCA1 sequence variants were identified among 122 Polish women with positive, in most cases moderate family history of breast and/or ovarian cancer, 80 controls and 34 unselected breast cancer tissue specimens. All mutations and variants were germline. The 4153 delA frameshift mutation, the Tyr105Cys missense mutation and two cases of the alteration in intron-20 were found in the group of healthy women with positive family history. Two other cases of the intronic insertion were found in unselected controls. Their carriers had no family history of breast or ovarian cancer but other cancers occurred in their families. The 1782 Trp/STOP nonsense mutation and one case of the insertion in intron-20 were first found in tissue specimens of breast cancer patient and breast/ovarian cancer patient, respectively. Their carriers also had no family history of breast or ovarian cancer. The distribution of the insertion in intron-20 in analysed groups and results of RT-PCR experiments suggest a less prominent role for this variant considered earlier a splicing mutation. This study shows also, that more population-oriented research is needed, involving women with less profound or even no family history of breast and ovarian cancer, to better understand the role and significance of different BRCA1 variants and mutations.

[Two cases of cancer developing from endometriosis in the minor pelvis 5 and 2.5 years after hysterectomy and adnexectomy]. / Dwa przypadki raka rozwijajacego sie na podlozu gruczolistosci w miednicy mniejszej po 5 i 2,5 latach po wycieciu macicy z przydatkami.

In two cases, 5 and 2.5 years after hysterectomy and bilateral adnexectomy (myomata uteri, bilateral endometrioid cysts and pelvic endometriosis) endometrioid carcinoma developed. Finding out the endometriosis during operation is an indication for precise removing of its residual and following careful examinations over the period of many years.

Results of topical treatment of HPV infection in the uterine cervix using interferon beta, 13-cis-retinoic acid and TFX.

Local treatment was administrated to 130 women, in whom colposcopy, cytology and (if the latter was abnormal) histology pointed to HPV infection in uterine cervix. The best results were obtained with 13-cis-retinoic acid (provitamin A) in conjunction with interferon beta (94.2% cured patients), followed by interferon beta in combination with TFX (90% cured patients) and interferon beta administered alone (88% cured patients).

Affinity electrophoresis for diagnosis of cancer and inflammatory conditions.

Affinity electrophoresis, with concanavalin A and Lens culinaris agglutinin as ligands, was applied to study the microheterogeneity of serum proteins, with special emphasis on alpha 1-fetoprotein and alpha 1-acid glycoprotein, two proteins of potential clinical value. A total of 602 samples of serum from patients with various neoplastic and inflammatory conditions were evaluated. Affinity electrophoresis provided useful information for differential diagnosis of cancers of various origins including hepatomas, metastatic liver cancers and yolk sac tumors. The method also proved indispensable for the detection of intercurrent infection in patients with rheumatoid arthritis, systemic lupus erythematosus and scaled burns.

Microheterogeneity of human alphafetoprotein.

The data available so far on the microheterogeneity of human AFP are reviewed. Molecular and developmental bases for the microheterogeneity of human AFP are discussed. Special attention is paid to the application of the studies on AFP microheterogeneity for the prenatal diagnosis of developmental malformations and for the differential diagnosis of some cancers. The results obtained using different methods are compared with special reference to lectin affinity electrophoresis and lectin affinity chromatography. In addition, a self-explanatory nomenclature of AFP variants is proposed in order to simplify the comparison of results obtained by different investigators. This has already been accepted by several investigators. The eventual simplification of methods applicable for clinical purposes and especially the difficulties encountered in experiments with monoclonal antibodies are also discussed.

Detection of messenger RNAs of alpha-fetoprotein and albumin in a human hepatoma cell line by in situ hybridization.

Detection of RNA transcripts within individual cells by in situ hybridization provides a powerful means for identifying specific cell types actively transcribing specific genes. We have applied this technique in order to analyze expression of the alpha-fetoprotein and albumin genes in a human hepatoma cell line, HuH-7. Using either 3H- or 35S-labeled human alpha-fetoprotein complementary DNA clone as a probe, we found that essentially all HuH-7 cells contained alpha-fetoprotein mRNA, although in various amounts. This correlated well with the presence of alpha-fetoprotein in all cells as detected by the peroxidase-antiperoxidase immunoenzyme method. The intracellular concentration of albumin, on the other hand, was below the level of detection by the peroxidase-antiperoxidase method. Consistent with this observation, we could not detect albumin mRNA with 3H-labeled albumin complementary DNA probes. However, the use of 35S-labeled probes having higher specific activities and higher efficiency of grain development resulted in the detection of albumin mRNA in a small percentage of HuH-7 cells. A variety of parameters involved in the in situ hybridization technique was examined to establish conditions suitable for demonstrating the presence of high- and low-copy numbers of mRNA in various cell and tissue preparations.

Microheterogeneity forms of alpha-fetoprotein present in amniotic fluid.

The microheterogeneity forms of human alpha-fetoprotein present in amniotic fluid were studied in affinity immunoelectrophoresis using two lectins: concanavalin A and Lens culinaris agglutinin. Ninety-seven samples of amniotic fluid collected throughout gestation were obtained from pregnancies in which there was no fetal malformation. Two samples of amniotic fluid from pregnancies complicated by anencephaly and one sample obtained from an empty gestational sac were also studied. The changes in relative concentrations of AFP microheterogeneity forms during gestation are described. It is suggested that concanavalin A is suitable for the demonstration of malformations of the central nervous system in the first and second trimester of pregnancy while in the third trimester it would be of no value. The results of this study suggest that Lens culinaris agglutinin might be suitable for this purpose in the third trimester of gestation, but this needs to be confirmed in a larger study. The optimal concentrations of both lectins were investigated. In addition, a simple method of rocket affinity electrophoresis is also described. The possible mechanisms responsible for the changes in relative concentrations of microheterogeneity forms of alpha-fetoprotein during gestation and in pathological conditions are discussed.

Microheterogeneity of alpha-fetoprotein in patient serum as demonstrated by lectin affino-electrophoresis.

Microheterogeneity of alpha-fetoprotein (AFP) present in the sera of 76 patients was studied by lectin affino-immunoelectrophoresis. Seventeen patients had benign liver disorders and the remaining 59 patients were treated for primary or secondary liver cancer or yolk sac tumour. By means of Con A, AFP was divided into two variants, while lentil lectin (LCA) made it possible to separate AFP in three variants. In some patients the relative concentrations of Con A and LCA AFP variants were similar; these patients were believed to produce AFP of the same 'profile'. Fourteen AFP profiles were observed by estimation of the area enclosed by precipitates corresponding to respective AFP variants. It was also possible to estimate the AFP profile on the basis of a simple visual analysis of the electrophoretic plates. The obtained results indicate that the AFP profiles of patients with cancer were variable. In spite of variations of the AFP profile in cancer patients, in most cases it was possible to differentiate primary liver cancer from yolk sac tumour and from liver metastases of cancer. In addition, in two-thirds of hepatoma patients the AFP profile was different from the profile observed in patients with benign liver disorders.

The in vitro production of alpha--fetoprotein variants by human fetal organs.

We have demonstrated the immunological identity of alpha-fetoprotein (AFP) produced in vitro by various fetal organs such as the yolk sac, liver, intestine and kidney. AFP produced in vitro by these organs, present in fetal serum and in samples of amniotic fluid was, however, heterogeneous when studied by lectin affinity immunoelectrophoresis. Results obtained with Concanavalin A were comparable to those obtained with Lentil agglutinin, but Concanavalin A separated AFP into two variants only, while Lentil agglutinin recognised three AFP variants. AFP produced by the yolk sac, intestine and kidney was similar in their proportions of variants and differed clearly from liver AFP. The ratios of AFP variants present in fetal serum were dissimilar to those of yolk sac AFP and of liver AFP. Relative amounts of AFP variants in fetal serum were similar in the first and last trimester of gestation. The proportions of AFP variants present in amniotic fluid in the first trimester of gestation were different from fetal serum, but, at the end of pregnancy, they become identical to those observed in fetal serum. Application of AFP variants for prenatal and cancer diagnosis is discussed.