RU2623857C1 - Method for precancerous changes identification in mammary gland - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: to detect precancerous changes in the mammary gland, interleukin 8 (IL-8) concentration is determineed in the blood cells sample supernatant incubated with a cancer-embryonic antigen (CEA) and without CEA. After that, the influence index (II) of the cancer-embryonic antigen (CEA II) on IL-8 production (CEA II IL-8) is calculated using the formula: CEA II IL-8 = A/B, where A is the level of cytokine production under the influence of CEA , B is the level of spontaneous cytokine production. At the CEA II IL-8 value not exceeding 14, precancerous changes in the mammary gland (sclerosing adenosis or mammary gland proliferation) are revealed.

EFFECT: application of this method allows to detect precancerous changes in the mammary gland, which allows timely initiation of adequate treatment.

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Description

The invention relates to medicine.

Known methods for identifying pathological changes in the mammary gland, which include:

1. Fine needle biopsy, which consists in taking a puncture from various areas of the mammary gland and studying the cellular composition of punctate. The result of a cytological study largely depends on how and from which sections of the mammary gland the material was taken and how it was processed. Fine needle biopsy in only 30-54% of cases gives an objective result associated with the above factors. Thus, in 46-70% of cases, a fine needle biopsy is not indicative (Jebsen PW, Hagmar V.M., Nesland JM Metaplastic breast carcinoma. A diagnostic problem in fine needle aspiration biopsy // Acta Cytol. 1991. Vol. 35. P. 396-401). In addition, a fine-needle biopsy causes microtrauma, which leads to accelerated tumor growth if there is a tumor (Zbitskaya M.A. The role of biopsy in the progression of breast cancer / M.A. Zbitskaya // Human health as the basis of national security: materials of the Siberian Congress. - Krasnoyarsk, 2006 .-- S. 311-313).

2. Tissue biopsy is to take a tissue sample from the breast when a tumor is suspected. A small piece is cut out for a more detailed study. After a thorough analysis, the pathologist concludes that the tumor is benign or malignant (Semiglazov VF Surgical treatment of breast cancer, history and modernity. Practical oncology. 2002. V. 3. No. 1. P. 21-28; Semiglazov B .F. Semiglazov VV Breast Cancer Screening. Practical Oncology. 2010. V. 11. No. 2. P. 60-65). This method is traumatic and technically difficult, and taking a piece of tumor tissue, especially repeatedly, activates tumor progression and can cause metastasis (Zbitskaya M.A. Effect of tumor biopsies on the progression of the process in patients with breast cancer / M.A. Zbitskaya, Uralsky medical journal - 2007. - No. 2. - S. 67-70).

3. If breast tumor pathology is suspected, the definition of cancer embryonic antigen (CEA), which is a glycoprotein with a molecular weight of 180 kDa, is most often used in blood or plasma. An increased concentration of CEA in the blood serum is detected in 30-50% of patients with breast cancer (A. Kishkun, Guidelines for laboratory diagnostic methods. - M., GEOTAR-Media, 2009. - 800 p.), I.e. this indicator is not sensitive enough (prototype).

However, all of the above methods do not allow to detect precancerous changes in the mammary gland, which include sclerosing adenosis, intraductal, intralobular proliferates, radial scar (Pathological anatomy National leadership / Ch. Ed. M.A. Pal'tsev, L.V. Kaktursky, O. V. Zairatyants. - M., GEOTAR Media, 2011 - 1264 p.).

Disclosure of invention

The proposed method for detecting precancerous changes in the mammary gland is characterized in that the concentration of interleukin 8 (IL-8) in the supernatant of a patient’s blood cell sample incubated with cancer-embryonic antigen (CEA) and without CEA is determined, and then the cancer impact index (IV) is calculated -embryonic antigen (IV CEA) for the production of IL-8 (IV CEA IL-8) according to the formula: IV CEA IL-8 = A / B, where A is the level of cytokine production under the influence of CEA, B is the level of spontaneous cytokine production, and with the value of IV REA IL-8 not exceeding 14, precancerous Changes in the mammary gland (sclerosing adenosis, or proliferative breast).

The present invention makes it possible to detect precancerous changes in the mammary gland, based on the determination of the cytokine-producing function of immunocompetent blood cells under the influence of cancer-embryonic antigen (CEA) on them. The method allows timely detection of precancerous changes and begin adequate treatment.

The proposed method is more sensitive and specific compared to the prototype. In the examined groups of patients, the proposed method showed 100% sensitivity and 100% specificity.

The proposed method is as follows. Whole blood taken from a patient's vein in an amount of 1 ml is placed in a sterile vial with 4 ml of medium (sterile DMEM growth medium) containing heparin 2.5 IU / ml and gentamicin 100 μg / ml. The contents of the vial are gently mixed, then 0.980 ml of diluted blood is taken from it under sterile conditions and placed in a sterile vial containing 0.020 ml of CEA (Canceroembryonic Antigen Human Colon Adenocarcinoma Cell Line) manufactured by Calbiochem, Germany, i.e. the concentration of CEA is 2 μg / ml . The contents of the vial are gently mixed and placed in a thermostat at 37 ° C for 24 hours. In parallel, under the same conditions, blood cells are incubated in the same nutrient medium without CEA (to determine the spontaneous production of cytokines). During the first hour of incubation, the contents of the vials are gently mixed 1 time. After incubation by centrifugation at 3000 rpm for 10 minutes, blood cells are precipitated and the supernatant is taken from a sample of cells incubated with CEA (production under the influence of CEA) and from a sample of cells incubated without CEA (spontaneous production), in which enzyme-linked immunosorbent assay determine the concentration of IL-8. IV REA IL-8 is calculated by the formula: IV REA = A / B, where A is the level of cytokine production under the influence of CEA, B is the level of spontaneous cytokine production. IV REA IL-8 is expressed in arbitrary units.

IV REA IL-8 was determined in 18 patients, in And of whom a benign tumor - fibroadenoma (FAM) was detected, in 7 - precancerous changes (in six - sclerosing adenosis and in one - intraductal proliferate).

Statistical analysis of the data was carried out according to the Mann-Whitney criterion, the median (Me), quartiles (Q _1; Q ₃ ) and the significance of the difference in p were determined.

In 11 patients with fibroadenoma, Me = 58.14 (Q ₁ = 27.14; Q ₃ = 91.39). All women in this group of IV REA IL-8 was above 14.00 conventional units.

In 7 women with sclerosing adenosis and intraductal proliferate related to precancerous lesions, Me = 8.66 (Q ₁ = 2.22; Q ₃ = 11.60). All women in this group of IV REA IL-8 was below 14.00 conventional units. The significance of differences in the RE of IL-8 between groups of women with FAM and with precancerous lesions (sclerosing adenosis and intraductal proliferate) was p = 0.0001, which reflects a high degree of difference in the performance of these groups.

In patients with fibroadenoma IV REA, IL-8 was in the range of 18.68-112.87 conventional units, in patients with precancerous changes (with sclerosing adenosis and intraductal proliferate) in the range of 2.03-13.83 conventional units.

The presence of precancerous changes and a benign breast tumor were monitored by a thorough histopathological examination of biopsy specimens.

Thus, according to the invention, in 100% of cases, precancerous changes are detected - sclerosing adenosis and intraductal proliferate.

Clinical examples

1. Patient B-a, 47 years old. The conclusion of the pathologist: sclerosing adenosis of the mammary gland. IV REA IL-8 - 6.59 conventional units.

2. Patient P-a, 54 years old. The conclusion of the pathologist: sclerosing adenosis of the mammary gland. IV REA IL-8 - 2.03 conventional units.

3. Patient Pa, 46 years old. Conclusion of the pathologist: intraductal proliferation of the mammary gland. IV REA IL-8 - 11.60 conventional units.

4. Patient K., 57 years old. Conclusion of the pathologist: breast fibroadenoma. IV REA IL-8 - 27.14 conventional units.

Claims (1)

A method for detecting precancerous changes in the mammary gland, characterized in that the concentration of interleukin 8 (IL-8) in the supernatant of a patient’s blood cell sample incubated with cancer-embryonic antigen (CEA) and without CEA is determined, and then the cancer impact index (IV) is calculated -embryonic antigen (IV CEA) for the production of IL-8 (IV CEA IL-8) according to the formula: IV CEA IL-8 = A / B, where A is the level of cytokine production under the influence of CEA, B is the level of spontaneous cytokine production and the value of IV CEA IL-8, not exceeding 14, detect precancerous changes in m the mammary gland (sclerosing adenosis or proliferation of the mammary gland).