Studies on Response to the Challenging Dose of X-rays in Lymphocytes of Prostate Cancer Patients and...
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Studies on Response to the Challenging Dose of X-rays in Lymphocytes of Prostate Cancer Patients and Healthy Donor. M. Krzysiek 1 , A. Cebulska-Wasilewska 1,2 , A. Panek 1 , W. Lipczy ń ski 3 , M. Dobrowolska 3 , Z. Dobrowolski 3

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M krzysiek 1 a cebulska wasilewska 1 2 a panek 1

Studies on Response to the Challenging Dose of X-rays in Lymphocytes of Prostate Cancer Patients and Healthy Donor

M. Krzysiek 1, A. Cebulska-Wasilewska 1,2, A. Panek 1,

W. Lipczyński 3, M. Dobrowolska3, Z. Dobrowolski 3

1Department of Radiation and Environmental Biology, INP PAN, Radzikowskiego 152, 31-342 Kraków, Poland

2Chair of Epidemiology and Preventive Medicine, Kopernika 7, Collegium Medicum Jagiellonian University, Kraków, Poland

3Department and Clinic of Urology CM UJ, Grzegórzecka 18, Kraków, Poland.


Aim of study

Aim of study

Aim of the study was to investigate variability beetwenprostate cancerpatients or benign prostate hyperplasia diseasepatients in a cellular response to the challenging dose of X-rays. The results were compared with the response of healthy donor’s cells.

It is a part of wider study which aim was to understand the mechanisms of cellular response that could help in deciding wheter prostate cancer patient should recieve radiotherapy or surgery.


Materials and methods

Materials and methods

Investigated groups:

  • Prostate Cancer (PC) patients: 31 males, avarage age:62.6y

  • Benign Prostate Hyperplasia (BPH)patients: 29 males, avarage age: 68.5y

  • Internal Standard cells (IS): healthy male donor

    DNA repair competence assay (fig.1):

  • Challenging dose of X-rays

  • DNA damage detection:alkaline version ofSingle Cell Gel Electrophoresis (SCGE)


Materials and methods1

Materials and methods

The analysis of DNA in the comets were applied with the use of two parameters from the Komet 3.0 or Komet 5.5 software (Kinetic Imaging Company, Liverpool, UK):

  • TDNA – tail-DNA (percentage of DNA in the comet tail)

  • TM – tail moment (which expresses the percentage ofDNA in the tail multiplied by the tail length TL—the length of the comet tail measured from the edge of the comet head)


M krzysiek 1 a cebulska wasilewska 1 2 a panek 1

Patient’s lymphocytes

Internal standard lymphocytes

Viability test

X-irradiation on ice, total dose 3Gy

Radiosensitivity was evaluated as follows:

ST-DNA = T-DNAx – T-DNA0

T-DNAx– measured after irradiation

T-DNA0 - measured without irradiation

Degree of DNA damage not repaired during the incubation (Residul Damage):

RDT-DNA=T-DNA40min/T-DNAx100%

T-DNA40min – measured after 40 min incubation

Repair: 40 min, 37oC, medium: 80%RPMI,20%FCS

Comet assay

Figure 1. Sheme of repair competence assay


M krzysiek 1 a cebulska wasilewska 1 2 a panek 1

Results

Table 1 The avarage levels of Radiosensitivity and Residual Damage evaluated for investigated groups of patients standardized with IS cells.

Significant (p<.05) increase in RDTM in cells of BPH patients and PC patients in comparison with Healthy Donor cells.

Significant increase in RDT-DNA in cells of BPH patients in comparision with PC patients

Significantly (p<.05) increased Radiosensitivity of lymphocytes collected from PC and BPH patients in comparison with IS cells

Table 2 The avarage levels of Radiosensitivity and Residual Damage evaluated for healthy person (IS)


M krzysiek 1 a cebulska wasilewska 1 2 a panek 1

Variability of celluar radiosensitivity

ST-DNA = 27.4

 SD = 5.0

Figure 1 Celluar radiosensitivity of PC patients (ST-DNA) described as the dispersion from avarage of healthy donor cells.

In a group of PC patients the range of Radiosensitivity

(ST-DNA)was from 16.9 to 40.9, while from 18.7 to 36.9 in group of BPH patients

ST-DNA = 27.5

 SD = 5.3

Figure 2 Celluar radiosensitivity of BPH patients (ST-DNA) described as the dispersion from avarage of helathy donor cells.


M krzysiek 1 a cebulska wasilewska 1 2 a panek 1

Variability of Residual Damage

RDT-DNA = 55.1

 SD = 16.9

Higher variability of RDin a group of PC patients than for BPH patients

Figure 3Residual damage (RDT-DNA) of PC patients evaluated as the dispersion from avarage of healthy donor cells.

In a group of PC patients the ratio between highiest and the lowest RDTM was 6.2.

In a group of BPH patients the ratio was 5.6

ST-DNA = 64.7

 SD = 12.6

Figure 4Residual damage (RDT-DNA) of BPH patients evaluated as the dispersion from avarage of healthy donor cells.


Conclusion

Conclusion

  • Significant increase in Residual Damage in cells of BPH patients and PC patients in comparison with Healthy Donor cells.

  • Significantly increased Radiosensitivity of lymphocytes collected from PC and BPH patients in comparison with Healthy Donor cells

  • Higher variability of Residual Damagein a group of PC patients than for BPH patients

    It is hopefully possible to identify a subpopulations of patients whose cells are more radiosensitive or resistant or express higher or lower efficiency of DNA damage repair. In a consequence it might be useful information in a process of optimization and individualization of the therapy.


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