SU1509040A1 - Method of diagnosis of the lung cancer - Google Patents

Abstract

The invention relates to immunoradiological studies and can be used as a diagnostic test for examining patients with suspected lung cancer. The purpose of the invention is to simplify and improve the accuracy of diagnosis. The cytotoxic activity of platelets is immunologically determined. The peripheral blood platelets of patients with lung cancer have a specific cytotoxic effect against ACL cells of the transplantable lung cancer line, with a diagnostic accuracy of 89.5%, whereas platelets of patients with chronic inflammatory lung diseases and platelets of healthy individuals are not capable of carrying out such a lytic effect, . The specificity of the method is 100%. In the prototype, the specificity of the method is 71.7%, and the diagnostic accuracy is 88.7%. Technically, the proposed method is simpler to perform and less traumatic for the subject. Thus, the proposed method for the diagnosis of lung cancer can be proposed as one of the diagnostic tests for a comprehensive examination of patients with suspected lung cancer. 2 tab.

Description

The invention relates to immunoradiologic.m studies and can be used as a diagnostic test for examining patients with suspected lung cancer.

The purpose of the invention is to simplify the method and improve the accuracy of diagnosis of lung cancer.

Example. 10 ml of the patient's venous blood are introduced into a 20 cm tube with 5% Na2 EDTA solution. The ratio of blood and 1 a2EDTA is 9: 1. The tube is allowed to sit at room temperature for 1 hour to sediment erythrocytes. The tube is then centrifuged for 10 minutes at 200 g, about 4/5 of the plasma with platelets is taken from the supernatant and transferred to another test tube, which is centrifuged for 10 minutes at 200 g. The supernatant containing only platelets is transferred to a plastic tube and centrifuged for 15 minutes at 1600 g. The platelet pellet is resuspended in 1 ml of RPM1 1640 medium (G1BCO) containing 5% fetal bovine serum, 1% L-glutamine, 1% HEPES buffer and 100 units / ml penicillin and streptomycin (medium A). The number of platelets was counted in a Gorev chamber using microscopy, after which the platelet suspension was diluted with medium A to a concentration of 100 x 10 / ml. Using serial dilutions, suspensions with platelet concentrations of 50-10, 2510 12.10 10, 5-10, 2.5-10, 1.2-10, 0.610 0.6-10 0.3-10 N 0.15 are prepared. -Kf / ml.

FROM

About About 4

As the target cells (KM), a monolayer culture of a transplantable human lung adenocarcinoma (ACL) strain is used.

In order to obtain labeled ° Cg CM, on the eve of the experiment, b-Yu ACL cells are incubated in 0.5 ml of medium 199 enriched with 10% bovine serum (medium B), containing 75-100 µCi of Na2 Cr04, periodically shaking the tube. After 40-60 minutes of incubation, the cells are washed three times in medium B for 10 minutes at 200 g and seeded with 2X10 cells in 0.2 ml of medium A into flat-bottomed 96-well microplates. The next day, before adding the platelets, the CM is washed twice with medium B. The platelets in 0.2 ml of medium A from each dilution are added to the CM, and the following platelet ratios are obtained: KM 1000: 1, 500: 1, 250: 1, 125: 1, 100: 1, 50: 1, 25: 1, 12: 1, 6: 1, 3: 1 and 1.5: 1. After 18 hours of incubation at 37 ° C in an atmosphere of 5% COO, all liquid is withdrawn from each well to determine the radioactivity in a y-spectrometer. The percentage lizirovapnyh KM evaluated by the formula:

% lysis KM 11 X100%,

where about (experience) is the output of the isotope from

lysed CM;

with (spontaneous release - release of isodes) top intact

KM;

n (full inclusion, complete inclusion) of the isotope in the CM.

The percentage of cytolysis can also be determined by the method of staining cells with trypan blue, but this method is used only for suspension cultures and is not acceptable for single-layer culture of AKL cells.

Using the indicated formula for calculating% of cytolysis and substituting values for the number of pulses per minute, the percentage of lysed CM in each ratio is determined (Table 1).

In tab. Figure 1 shows the cytolytic activity of platelets of a patient with lung cancer against AKL cells (growth of effector: KM ratios).

Thus, in the example above, firstly, the maximum level of cytolysis of CM (29.8%) is noted when the effector ratio of CM is 50: 1. This ratio is considered optimal and is used in all further experiments; secondly, 0 level of cytolysis exceeds the discriminatory value by 19.8%. This makes it possible to diagnose lung cancer in this patient, which was later confirmed by a morphological study of the bioptag of the tumor.

In tab. 2 compares the cytotoxicity of peripheral blood platelets in patients with lung cancer and those examined in the control group against ACL cells.

As a result of the conducted studies, it was found that the method for determining the cytotoxicity of peripheral blood platelets examined against ACL cells is technically simpler to perform, does not require complicated and expensive equipment for bronchological research and bronchoalveolar lavage.

The study of the cytotoxic activity of platelets as a diagnostic test for lung cancer is a novelty of the invention.

Claims (1)

Isorethorne formula A method for diagnosing lung cancer by radioisotope research in biological fluids, characterized in that, 5 in order to simplify and improve the accuracy of the method, the study is carried out with blood platelets, adding them to the labeled cells of the interrupted lung adenocarcinoma line in a ratio of 25-75 ): 1, incubated, determine the percentage of lysed 0 cells and when the value of this indicator more or equal diagnose lung cancer. 1000: 1500: 1 3,82,5. Table 1 23.2 ± 8.9 4 3 + 2 7 h-, j -, / Table 2 nineteen , 001 21