Hec1/Nek2 Mitotic Pathway Inhibitor INH1 Inhibits the Cell Kinetic Parameters of A549 and HeLa Cells

Abstract In this current study, antiproliferative effect of Hec1/Nek2 Mitotic Pathway inhibitor INH1 was investigated in adenocarcinomic human alveolar basal epithelial cells A549 and human cervix carcinoma HeLa cell lines in vitro. To this end cell index values by xCELLigence Real‐Time Cell Analysis DP instrument, mitotic index, BrdU proliferation assay and apoptotic index analysis were used. The results of the current study showed that INH1 had cytostatic and cytoskeletal effects on A549 and cytostatic effect on HeLa cells. The IC50 concentration was determined as 56 µM with the xCelligence device for both cell lines. IC50 concentration was used for all other parameters. While this concentration decreased the mitotic index BrdU proliferation values, it increased the apoptotic index values of both of cell lines. There were significant differences between the control and the experimental groups (p<0.05). The results of the present study suggest that INH1 may serve as a promising treatment option for different types of cancer.

The effect of aging on micronuclei frequency and proliferation in human peripheral blood lymphocytes.

INTRODUCTION: Increase in the instability of cellular genome with an increasing age is the result of an accumulation of cellular damage and mutations. This instability which might be observed as chromosome damage or chromosome losses can be measured by the micronucleus technique. AIM: The aim of this study was to investigate the effect of aging and oxidative stress induced by non-toxic levels of H2O2 on micronuclei induction and their relationship to cell proliferation in human peripheral blood lymphocytes. MATERIALS AND METHODS: Healthy volunteers with different ages were choosen. Spontaneous and H2O2 induced micronuclei frequencies were measured in peripheral blood lymphocytes of 30 volunteers by the micronucleus method. RESULTS: Spontaneous micronuclei frequencies increased first then started to decrease after 50 years of age. This biphasic response was significantly higher than micronucleus (MN) frequencies induced by H2O2 (P < 0.05), which followed the similar shape of response to increasing ages with lower frequencies. Proliferative capacity of cells either treated with H2O2 or not did not differ with an increasing age giving similar responses. CONCLUSION: These results indicate biphasic character of chromosome damage; first increase and decrease after 50 years with an increasing age. But this change pattern was not correlated with the steady state of proliferation capacity of cells through an increasing age. Decreases in H2O2-induced MN frequencies compared to spontaneous MN frequencies may be inducing an apoptosis by H2O2 treatment leading to underscoring damaged cells.

Clinical significance of silver binding nucleolar organizer regions(AgNORs) in transitional cell carcinoma of the bladder : The correlative study with flow cytometric SPF, PI and PCNA / 대한비뇨기과학회지

Nucleolar organizer regions(NORs) contain coding genes for ribosomal RNA and contribute the regulation of cellular protein synthesis. AgNORs numbers correlate with growth fraction and have been reported the AgNORs counts may have a diagnostic and prognostic utility in other human tumors. We investigated the diagnostic usefulness of AgNORs staining technique as a discriminant for malignancy and assessed the value as a potential method for the estimation of cell kinetics. In addition. we compared the AgNOR counts with flow cytometric analysis of ploidy, S-phase fraction, proliferation index, and PCNA expression rate. There was a statistically significant difference of AgNORs counts between superficial bladder tumor and invasive bladder tumor. But there was no relationship between the mean number of AgNORs per nucleus and histological grade. DNA aneuploid group was associated with higher AgNORs counts than diploid group, but the difference was statistically insignificant. The mean number of AgNORs per nucleus had significant relationship to SPF(r=0.43, p<0.05) and PI(r=0.41, p<0.05.) We concluded that this method alone does not offer a reliable histological discriminant for malignancy. Further studies are needed to confirm that AgNORs counting is a useful method for evaluating the proliferative activity and this technique may serve as a prognostic factor additional to the current histopathological grading criteria of the bladder cancer.

The clinical significance of expression of proliferating cell nuclear antigen in transitional cell carcinoma of the bladder : The comparative study with histopathological grade and clinical stage / 대한비뇨기과학회지

The expression of proliferating cell nuclear antigen (PCNA) was assessed immunohistochemically in 47 cases of transitional cell carcinoma of the bladder and 3 cases of normal bladder with anti- PCNA/cyclin monoclonal antibody, using routinely processed tissue sections without interferring with histopathological diagnosis. The PCNA expression rates were compared with Ash histologic grade and clinical stage. In bladder cancer, the PCNA expression rate ranged from 3.8% to 32.7 % (mean value 11.2 %). Bladder cancer with Ash grade IV showed the highest PCNA expression rate (mean value 15.8 % ) and cancer with Ash grade I showed the lowest PCNA expression rate (mean value 8.3%). There were statistically significant differences of PCNA expression rates according to Ash grades (P=0.02. Kruskal-Wellis test). When clinical stage was analyzed to assess the relationship to PCNA expression rate invasive bladder cancers were associated ith higher PCNA expression rate then superficial bladder cancer (mean value of stage A; 8.7 %, stage B and C; 16.5 %). and the difference was statistically significant (P=0.003. Kruskal-Wallis test). Also, there was positive linear relationship between PCNA expression rate and Ash grade with regression analysis (r=0.573, P<0.0001, Y=4.41X +0.79). These results suggest that PCNA is useful as a unclear antigenic marker of cellular proliferation and offers an opportunity for analyzing cell kinetics successfully in formalin-fixed, paraffin-embeded tissue sections of transitional cell carcinoma of the bladder. It will be merited as a simple and powerful method to detect transitional cell carcinoma of the bladder with high potential of invasion, metastasis and clinical progression.

Comparison of brain tumor growth kinetics by proliferating cell nuclear antigen (PCNA) and bromodeoxyuridine (BrdU) labeling

The bromodeoxyuridine (BrdU) labeling study provides valuable cell kinetic information for individual tumors that could suggest the prognosis of each patient who had a tumor. Recently, a monoclonal antibody against the proliferating cell nuclear antigen (PCNA or cyclin), a nuclear protein expressed in proliferating cells, was developed which could be used on formalin fixed, paraffin embedded tissue. The purpose of this study was to compare the cell kinetic data obtained by the BrdU labeling study and the PCNA method in the same patient. The relationship between labeling indices of BrdU incorporated into S-phase and PCNA expressed by cycling cells was investigated in 31 patients with brain tumors. Both of the labeling indices showed good correlation with histological grade of the tumor. The values of the PCNA labeling index (LI) were parallel but higher than those of the BrdU LI, and the relation PCNA LI = 2.2 x BrdU LI + 0.8 (r2 = 0.86) was obtained. The results of this study show that PCNA could replace the BrdU method for identifying the proliferating cells, and the major advantages of PCNA method is that it could be done without any pretreatment and avoid injection of the teratogenic agent for diagnostic purpose.

The clinical significance of expression of proliferating cell nuclear antigen(PCNA) in prostatic carcinoma: the correlative study with glandular differentiation and flow cytometric DNA analysis / 대한비뇨기과학회지

The expression of proliferating cell nuclear antigen (PCNA) was assessed immunohistochemically in I8 patients with benign glandular hyperplasia (BPH) and 40 patients with prostatic carcinoma with anti-PCNA, cyclin monoclonal antibody. using routinely processed tissue sections without interfering with histopathological diagnosis. The PCNA expression rates were compared with flow cytometric DNA analysis in 29 patients and Gleason histologic grade in 40 patients with prostatic carcinoma. In BPH tissues basal cells show positivities for PCNA and secretory cells show essential positive reaction with anti-PCNA/cyclin monoclonal antibody. In prostatic carcinoma the PCNA expression rate ranged from 3.70% to 48. 53% (mean value 22.36%1. Prostatic carcinoma with Gleason grade 5 showed the highest PCNA expression rate (mean value 32.22%) and carcinoma with Gleason grade l showed the lowest PCNA expression rate (mean value 3.TS%). There were statistically significant differences of PCNA expression rate according to Gleason grades (p=0.001. Kruskal-Wallis test). When DNA ploidy was analyzed to assess the relationship to other tumor variables DNA aneuploid group was associated with higher Gleason grade and higher PCNA expression rate than diploid group. but the differences were statistically insignificant. The DNA aneuploid group had higher percentage of S-phase cells (S-phase Fraction) by flow cytometry than diploid group (mean value l3.48 =6 in diploid group and 19.78% in aneuploid group and this difference was statistically significant (p=0.01. Mann-Whitney test). From the above results. it is clear that PCNA expression is useful as an nuclear antigenic marker of cellular proliferation and offers an opportunity for analyzing cell kinetics successfully in prostatic carcinoma. It will be merited as a means to detect prostatic carcinomas with high potential for invasion. metastasis and clinical progression.

Flowcytometric Analysis of DNA Content and Cell Kinetics in Nervous System Neoplasms

No abstract available.

A Study on the Cell Kinetics of the Dysplastic Epithelium in the Stomach

This study was designed to evaluate the biological behavior of the dysplastic lesion of the stomach by applying immunohistochemical method for bromodeoxyuridine (BrdUrd). The results obtained were as follows. 1) In most hyperplastic and dysplastic lesions, the proliferative cell zones, loci of BrdUrd-labelled cells, were found in the upper later of the mucosa, whereas they were confined to the neck zone in the normal gastric mucosa. 2) The labelling indices (LIs), percentages of BrdUrd-labelled cells, were 11.0% to 13.6% in the normal gastric mucosa, and were 14.3% to 17.9%, 16.4% to 19.2% and 17.4% to 20.7% in the simple hyperplasia, in the atypical hyperplasia and in the dysplasia, respectively. These findings suggested that proliferative potential in hyperplasia and dysplasia were greater than that in normal gastric mucosa, the higher the grade of dysplasia being, the greater the proliferative potentials.

Mathematical models for optimizing tumour radiotherapy. I: A simple two compartment cell-kinetic model for the unperturbed growth of transplantable tumours.

Tumour growth kinetics has been analysed on the basis of interactions between two compartments comprising the proliferating and non-proliferating cells. Starting from the differential equations of growth of the cell-populations in the two compartments and assuming the process of intercompartmental cell transfers to be linear, an analytic expression on the variation of growth-fraction with the age of the tumour is obtained. The restricted conditions on the cell-cycle time and cell-loss-rate, under which these differential equations lead to a Gompertzian growth of the tumour, are critically analysed. The formalism permits the estimation of some important cell-kinetic parameters, like growth-fraction or cell-loss-factor, from a knowledge of the tumour-growth curve, cell-cycle-time and a single measurement of the cell-loss-rate of the matured tumour, provided the tumour follows a Gompertzian growth. The validity of the model has been verified with the experimental data on 4 different transplantable murine tumour systems. Usefulness of the model has been demonstrated by making some interesting predictions regarding the radiation response of the tumours from the cell-kinetic parameters.