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UDC 616.61-006.04-018.53-076.5
INTRATUMORAL MAST CELLS BY RENAL CELL CARCINOMA: CLINICO-MORPHOLOGICAL CORRELATIONS
1 Altai State Medical University, Barnaul
2 Altai branch of N.N. Blokhin Russian Cancer Research Center, Barnaul
I.P. Bobrov1, T.M. Cherdantseva1, N.G. Kryuchkova1, A.V. Lepilov1, A.F. Lazarev2, A.M. Avdalyan2, V.V. Klimachev1, I.V. Klimachev1, M.N. Medyalets1, A.V. Kazartsev1
The results of the study presented in this article show the interrelation of intramural stromal mast cells in tumors with some of the most important prognostic clinico-anatomical factors in the development of renal cell carcinoma. The authors demonstrate the possibility of using the factor of the number of intramural stromal mast cells in the tumor as an additional one for prognostic evaluation of the development of the disease and metastasis of the tumor. The relationship between this factor and the 5-year postoperative survival of patients with renal cell carcinoma has been identified and proven.
Key words: intramural mast cells, renal cell carcinoma, oncology, risk factors.
Interest in studying the microenvironment of malignant tumor cells, including stromal in-tratumoral mast cells (MC), has been preserved for the last decades [1-6].
MCs are present in the normal and pathologically altered kidney [7-8]. There is a small number of works devoted to the study of the prognostic value of MC in renal cell carcinoma (RCC). According to Tune B. et al. [9], the content of intra-tumoral MCs was significantly higher by kidney cancer (KC) than in normal renal tissue, which, according to the authors' findings, is due to their involvement in tumor necrosis. At the same time, the authors did not find the relationship between the number of MCs in the tumor and the important factors in the prognosis of RCC: the clinical stage, the size of the tumor node, the Fuhrman gradation, but the number of MCs increased significantly in the clear cell carcinomas. The high number of intratumoral MCs in the clear-cell carcinomas is explained by the significant metabolic requirement of the cells of this histological variant of KC. Similar results were obtained by other authors [10]. At the same time, Guldur M.E. et al. [11] found significant interrelations between the number of MCs in the tumor and the stage, the Fuhrman gradation, the size and the histological type of tumor.
Thus, the studies devoted to prognostic value of the study of intratumoral stromal MCs by RCC are scarce and contradictory, and therefore this component of the micro-environment of the tumor needs further study.
The purpose of this work was to study intra-tumoral stromal MCs by RCC, to search for relationships with important clinical and anatomical factors of prognosis and post-operative survival of patients.
Materials and methods
The operating material of 63 patients with RCC was studied. The average age of the patients was
58.2±1.2 years. There were 34 men (54%), women - 29 (46%). In all observations, RCC was classified in accordance with the WHO classification of 2004. According to the histological structure of the tumor, the following were presented: bright cell carcinoma - 48; granular-cell cancer - 4; papillary cancer - 5; chromophobic cancer - 3, and spindle cell carcinoma (sarcomatoid) - 3.
The prevalence of the tumor process was assessed according to the TNM classification of malignant tumors (edited by Sobin L.H., 2002) [12]. Among the patients, 47 (74.6%) observations corresponded to I clinical stage (T1N0M0); II stage (T2N0M0) - 3 (4.8%) observations; III stage (T1N1M0, T2N1M0, T3N0M0, T3N1M0) - 8 (12.7%) and IV stage (T4N0M0, T4N1M0, T2N1M0, T1N1M0, T3N1M0) - 5 (7.9%). The degree of malignancy of tumor cells was assessed by Fuhrman S.A. et al. (1982) [13]. 31 tumors (49.2%) corresponded to the degree of anaplasia G1, degree of anaplasia G2 - 14 (22.2%) tumors, the degree of anaplasia G3 -13 (20.6%), and the degree of anaplasia G4 - 5 (8%).
Carcinomas with regional and distant metastases constituted 11 (17.5%), localized tumors - 52 (82.5%).
The average size of the tumor node was 7.1 ± 0.3 cm.
Control was the tissue of the kidneys, taken from the most removed from the tumor sites, without pronounced phenomena of inflammation and sclerosis.
The material was fixed in a 10% neutral buffered formaline for 12-24 hours. Histological preparations were stained with hematoxylin and eosin, and for acid mucopolysaccharides with colloidal iron by Gal. TCs were detected by immunohisto-chemical method using monoclonal antibodies C-kit (CD117) (clone A4052, dilution 1: 200, DAKO) according to the manufacturer's recommended protocol. The average number of MCs was calculat-
ed in three fields of view with the zoom in the microscope x 400.
Information on the survival of patients was obtained from the register of malignant diseases and outpatient cards (for 2010-2016). The statistical processing of the material was carried out with the statistical package Statistica 10.0. In the normal distribution of data, the methods of parametric statistics (Student's t-test) were used to check statistical hypotheses, and if the obtained data did not meet the criteria of normal distribution (Shap-iro-Wilk criterion W = 0.89, p <0.01), then the Kolm-ogorov-Smirnov test or the Man-on-Whitney U-test were implied. The data were considered significant by p <0.05. The development of the survival curves was carried out according to the Kaplan-Meier method, the reliability of differences in survival rates was assessed using a log-rank test.
Results and discussion
The results of the study showed that in unchanged tissue, the kidneys of MCs were located singly or in groups. They had a round, oval or branched form and located mainly in the inter-
The mean value of intratumoral MCs in tumors in men was 2.6 ± 0.2, in women -2.4 ± 0.2 (p = 0.6).
Depending on the age of the patients, the mean MC numbers were as follows: in patients aged 3039 years - 1.0 ± 0.4; from 40 to 49 years - 3.7 ± 0.4; from 50 to 59 years - 2.5 ± 0.3; from 60 to 69 years - 2.4 ± 0.3, and from 70 to 79 years - 1.8 ± 0.25. Thus, the highest rates of intratumoral MCs in the tumor stroma were observed in the age group of patients from 40 to 49 years, and the lowest - in the group from 30 to 39 years.
By the analysis of the content of MCs in a tumor, depending on the clinical stage of the disease, the following results were obtained for TNM: in patients with the I clinical stage, the mean number of MCs was 1.85 ± 0.15; at stage II, the number of MCs increased to 2.7 ± 0.4 (p = 0.2); at stage III, the number of TC increased to 4.3 ± 0.4 (p = 0.003), and at stage IV, it was 6.3 ± 0.6 (p = 0.005).
stitial tissue, near the vessels and between the tubules. On average, the number of MCs in the kidney tissue was 2,3±0,2 in three fields of vision with the zoom of the microscope x 400.
Among all patients with RCC, irrespective of gender, age, histological type and other clinical and morphological parameters, the spread of mean values of the distribution density of TCs ranged from 0 to 10. The mean value of the distribution density of MCs by RCC was 2.5 ± 0.2 .
A comparison of the average number of intratu-moral MCs in the stroma of the tumor with prog-nostically important clinical and anatomic factors of RCC revealed some important correlations. MCs distribution density was correlated: with clinical stage of disease (r = 0,69; p = 0,0001), with tumor nodule size (r = 0,58; p = 0,0001), with the degree of anaplasia tumor by Fuhrman (r = 0,89; p = 0,001), with presence of regional and distant metastases (r = 0,48; p = 0,0001) and postoperative survival of patients (r = 0,40; p = 0,001). There were no interactions with the gender (r = - 0,03; p = 0,8), age (r = - 0,15; r = 0,25) and histological type of the tumor (r = 0,16; p = 0.19) (Table 1.).
Thus, at the III and IV stages there was a significant increase in MCs as compared with stage I, and at the IV stage, the number of MCs significantly increased in comparison with the III stage of the pathological process.
Depending on the histological variant of RCC, the highest average number of intratumoral MCs was found in spindle cell carcinoma (6.3 ± 0.6), and the lowest value was papillary cancer - 1.05 ± 0.4. The intermediate position was occupied by the bright cell carcinoma (2,2 ± 0,2) and chro-mophobic (2,4 ± 0,4) cancer and a granular cell variant of the bright cell carcinoma (6.1 ± 0.8). Thus, the greatest number of MCs was detected in the spindle cell variant of RCC, and the smallest - in the papillary version.
When comparing the degree of nuclear atypia according to Fuhrman with the number of intra-tu-mor MCs, the following data were obtained: by GI,
Table 1 Correlation relationships of the distribution density of MCs with clinical and morphological parameters of RCC
Clinical and morphological parameter Correlation coefficient (r) Significance
Clinical stage 0,69 0,0001
Size of tumor node 0,58 0,0001
Degree of tumor anaplasia by Fuhrman 0,89 0,001
Presence of regional and distant metastases 0,48 0,0001
Postoperative survival of patients 0,40 0,001
Gender 0,03 -
Age - 0,15 -
Histological type of the tumor 0,16 -
the number of TCs was 1.1 ± 0.1, by GII it increased to 2.0 ± 0.2 (p = 0, 000006), by GIII it was 4.8 ± 0.3 (p = 0.0000001) and by GIV it was 6.3 ± 0.5 (p = 0.007). Thus, a significant increase in the number of intra-tumoral stromal MCs was found with an increase in the degree of anaplasia of tumor cells.
With a tumor node size <7.0 cm, the number of stromal MCs was equal to 1.9 ± 0.15, and at a size
of > 7.0 cm the amount of MCs in the tumor was significantly increased to 4.7 ± 0.4 (p = 0, 0000001).
In tumors of patients without regional and distant metastases, the MC content was 2.0 ± 1.2, and in the presence of metastases it increased significantly to 4.7 ± 0.4 (p = 0.0000001) (Table 2.) ( Figure 1, a, b).
Table 2
The content of intratumoral stromal MCs in the tumor, depending on the clinical and morphological parameters of RCC
Clinical and morphological parameter Correlation coefficient (r) Significance (p)
Gender: male 2,6±0,2
female 2,4±0,2 -
Age: 30-39 years 1,0±0,4
40-49 years 3,7±0,4 p = 0,005
50-59 years 2,5±0,3 -
60-69 years 2,4±0,3 -
70-79 years 1,8±0,25 -
TNM stage: I stage 1,85±0,15
II stage 2,7±0,4 -
III stage 4,3±0,4 p = 0,003
IV stage 6,3± 0,6 p = 0,005
Histological variant of tumor:
papillary 1,05±0,4
bright-cell 2,2±0,2 p = 0,01
chromophobic 2,4±0,4 -
granular cell 6,1±0,8 -
spindle cell 6,3±0,6 -
Fuhrman gradation: GI 1,1±0,1
GII 2,0±0,2 p = 0,000006
GIII 4,8±0,3 p = 0,0000001
GIV 6,3±0,5 p = 0,007
Tumor node size: < 7,0 cm 1,9±0,15
> 7,0 cm 4,7±0,4 p = 0,0000001
Metastases: N0 2,0±1,2
N+ 4,7±0,4 p = 0,0000001
Figure 1. Density distribution of intratumoral mast cells in the stroma of kidney cancer, depending on the presence of metastases: a - localized tumor; b - tumor with metastases. Immunohistochemical staining on CD117. Zoom x 400
To assess the long-term results of 5-year postoperative survival of patients depending on the density distribution intratumoral stromal MCs, tumors were divided into 2 groups. 1st group constituted carcinomas with small number of MCs (<3), the 2nd group included carcinomas with large number of MCs (> 3). In the analysis of survival by Kaplan-Meier it has been found that when
the number of MCs in the tumor was <3, the cumulative proportion of survived to 1800th day was 1 (100%), and when the MCs quantity in tumors was > 3, the cumulative proportion of survived to 1800th day was down to 0.62 (62%). When comparing the survival rates in groups of patients with the use of log-rank test it was found that the data were significant (log-rank p = 0,00015) (Figure 2).
Figure 2. Cumulative proportion of 5-year postoperative survival of patients with RCC depending on the number
of intratumoral MCs in the tumor
Thus, the results of the study showed that the number of intratumoral stromal MCs in the tumor was interrelated with a number of important prognostic clinical and anatomic factors of RCC, and therefore this parameter can be used as an additional factor in predicting the course of the disease. An increase in the amount of intra-tumoral MCs in the tumor can be considered as a criterion of the likely occurrence of tumor metastases. A relationship between the number of stro-mal MCs and the survival of patients was revealed. The high level of MCs content in the tumor is an unfavorable factor in the 5-year postoperative survival of patients with RCC.
References
1. Lazarev A.F., Bobrov I.P., Cherdantseva T.M., Klimachev V.V., Bryukhanov V.M., Avda-lyan A.M., Lubennikov V.A., Hervald V.Ya. Mast cells and tumor growth. Siberian journal of oncology. 2011; 4: 59-63.
2. Cherdantseva T.M., Bobrov I.P., Klimachev V.V., Bryukhanov V.M., Lazarev A.F., Avda-lyan A.M., Hervald V.Ya. Mast cells in kidney cancer: clinical-co-morphological parallels. Medicine in Kuzbass. 2011; 2: 48-51.
3. Conti P, Castellani ML, Kempuraj D. et al. Role of mast cells in tumor growth. Ann Clin Lab Sci. 2007; 4: 315-322.
4. Maltby S, Khazaie K, McNagny KM. Mast cells in tumor growth: angiogenesis, tissue remodelling and immunemodulation. Biochim Biophys Acta. 2009; 1: 19-26.
5. Liu J, Zhang Y, Zhao J, et al. Mast cell: insight into remodeling a tumor microenvironment. Cancer Metastasis Rev. 2011; 2: 177-184.
6. Rigoni A, Colombo MP, Pucillo C. The role of mast cells in molding the tumor microenvironment. Cancer Microenviron. 2015; 3: 167-176.
7. Roberts IS, Brenchley PE. Mast cells: the forgotten cells of renal fibrosis. J Clin Pathol. 2000; 11: 858-862.
8. Ehara T, Shigematsu H. Mast cell in kidney. Nephrology. 2003; 8: 130-138.
9. Tuna B, Yorukoglu K, Unlu M, Mungan MU, Kirkali Z. Association of mast cells with mi-crovessel density in renal cell carcinomas. Eur Urol. 2006; 3: 530-534.
10. Mohseni MG, Mohammadi A, Heshmat AS, et al. The lack of correlation between mast cells and microvessel density with pathologic feature of renal cell carcinoma. Int Urol Nephrol. 2010; 1: 109-112.
11. Guldur ME, Kocarslan S, Ozardali HI, et al. The relationship of mast cells and angiogenesis with prognosis in renal cell carcinoma. J Pak Med Assoc. 2014; 3: 300-303.
12. Sobin LH, Wittekind Ch. UICC - TNM Classification of malignant tumors. New York: Wi-ley-Liss; 2002: 193-195.
13. Fuhrman SA, Lasky LC, Limas C. Prognostic significance of morfologic parameters in renal cell carcinoma. Am J Surg Pathol. 1982; 6: 655-663.
Contacts:
Corresponding author: Bobrov Igor Petrovich, Senior researcher of the Morphological Laboratory of the Center for Medical and Biological Research of the Altai State Medical University, Barnaul. Tel.: (3852) 669927. E-mail: science@agmu.ru
Cherdantseva Tatyana Mikhailovna, Doctor of Medical Sciences, Professor of the Department of Oncology, Radiation Therapy and Radiation Diagnostics with the course of FVE of the Altai State Medical University, Barnaul. 656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
Kryuchkova Natalia Gennadyevna, Assistant Professor of the Department of forensic medicine and pathological anatomy named after professor VN. Kryukov with the course of FVE of the Altai State Medical University, Barnaul. 656038, Barnaul, Lenina Prospekt, 40. Tel.: (3852) 408439. Email: science@agmu.ru
Lepilov Aleksandr Vasilyevich, Doctor of Medical Sciences, Head of the Department of forensic medicine and pathological anatomy named after professor VN. Kryukov with the course of FVE of the Altai State Medical University, Barnaul. 656038, Barnaul, Lenina Prospekt, 40. Tel.: (3852) 408439. Email: science@agmu.ru
Lazarev Aleksandr Fedorovich, Doctor of Medical Sciences, Professor, Head of the Department of Oncology, Radiation Therapy and Radiation Diagnos-
tics with the course of FVE of the Altai State Medical University, Barnaul. 656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
Avdalyan Ashot Meruzhanovich, Doctor of Medical Sciences, senior researcher of Altai branch of N.N. Blokhin Russian Cancer Research Center, Barnaul.
656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
Klimachev Vladimir Vasilyevich, Doctor of Medical Sciences, Professor of the Department of forensic medicine and pathological anatomy named after professor VN. Kryukov with the course of FVE of the Altai State Medical University, Barnaul. 656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
Klimachev Ilya Vladimirovich, resident of the Department of forensic medicine and pathological anatomy named after professor VN. Kryukov with the course of FVE of the Altai State Medical University, Barnaul.
656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
Myadelets Dmitry Nikolayevich, assistant
of the Department of Urology and Andrology with
courses of specialized surgery of the Altai State
Medical University, Barnaul.
656056, Barnaul, Komsomolsky Porspekt, 73.
Tel.: (3852) 364737.
Email: dmyadelec@yandex.ru
Kazartsev Artem Vladimirovich, assistant of the Department of forensic medicine and pathological anatomy named after professor VN. Kryukov with the course of FVE of the Altai State Medical University, Barnaul.
656045, Barnaul, Zmeinogorsky Trakt, 110. Tel.: (3852) 632620. Email: science@agmu.ru
