Method of determination Histological examination of a biopsy specimen according to the histological classification of the World Health Organization (WHO) with hematoxylin-eosin staining. IHC study using a spectrum of tissue-specific and prognostic antibodies (up to five antibodies) (peroxidase and avidin-biotin methods).

Study material See description

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Comprehensive study of biopsy specimens of metastatic formations of various localizations, including morphological description and assessment of the expression of the spectrum of tissue markers, to determine the histogenesis of the primary tumor focus.

According to statistics, in 3-15% of cases, malignant tumors manifest as metastases. Metastases without an identified primary focus are characterized by random, atypical localization and rapid progression of the process. The average life expectancy of patients with three or more metastatic lesions is three months. At the same time, the localization of the primary focus in 60-70% of patients is revealed only at autopsy. Timely IHC-diagnostics determines the tactics of treatment before identifying the primary focus.

Pathomorphological diagnostics of metastases of undetected tumors provides for the determination of their morphological type, clarification of the probable source of metastasis, assessment of the malignant potential. The first task is solved during the routine histological examination of biopsy material (staining with hematoxylin-eosin), the second - using special research methods: histochemical, immunohistochemical (IHC) or molecular genetic method FISH (Fluorescence In Situ Hybridization).

In routine morphological diagnostics, metastatic tumors, according to the recommendations of the European Society for Medical Oncology (ESMO) (2004), are divided into five large categories: adenocarcinoma, squamous cell carcinoma, neuroendocrine carcinoma, undifferentiated cancer, undifferentiated tumor. These morphological categories, along with data on the prevalence of the process, in many cases allow you to determine an adequate plan of examination (and treatment). For example, when detecting metastasis of squamous cell carcinoma in the cervical lymph nodes, an endoscopic examination of the upper respiratory and digestive tract is necessary. The IHC study, depending on the morphological type of the neoplasm, makes it possible to clarify the histogenesis of the tumor and / or to determine the probable localization of the primary focus.

For the IHC study of primary tumors and their metastases, a wide range of tissue-specific markers is used: -cyte-specific (clusters of leukocyte differentiation (CD - Clusters of Differentiation), smooth muscle actin, myoglobin, thyroglobulin); - markers of proliferation (Ki67, PCNA - Proliferating Cell Nuclear Antigen); -tumor markers - oncofetal antigens (fetoprotein, carcinoembryonic antigen); - hormones (estrogen, progesterone); -enzymes, protein products of cellular oncogenes, etc.

Unified algorithms for IHC study of tumor metastases without a specified primary focus have not been developed. Correct determination of the direction of differentiation of tumor cells and a number of biological parameters of the tumor in itself is an indication for the appointment of certain therapy regimens. For example, the detection of metastases of adenocarcinoma in the lymph nodes of the axillary region may be an indication for a therapy similar to therapy for breast cancer of the corresponding stage. Detection of the expression of estrogen and progesterone receptors in such adenocarcinoma may become an indication for the appointment of antihormonal therapy, regardless of the presence of a detectable tumor node in the mammary gland. An example of an algorithm recommended for use in differential diagnostics.

Material for research: tumor biopsy, fixed in a paraffin block.

Literature

1. Blaszyk H., Hartmann A., Bjornsson J. Cancer of unknown primary: clinicopathologic correlations. APMIS. 2003; 111 (12): 1089-1094.
2. Dabbs D.J. Diagnostic Immunohistochemistry: Theranostic and Genomic Applications. Elsevier, 4-th Edition. 2013: 960.
3. Di Patre P.L., Carter D. Sternberg's Diagnostic Surgical Pathology Review. Wolters Kluver, 2nd Edition. 2015: 488.
4. Dietal M., Wittekind C., Bussolati G., von Winterfeld M. Pre-Analytics of Pathological Specimens in Oncology. Srringer. 2015: 133.
5. Dodd L.G, Bui M.M. Atlas of Soft Tissue and Bone Pathology. DemosMEDICAL. 2014: 720.
6. Edge S.B, Byrd D.R., Compton C.C., Fritz A.G., Greene F.L., Trotti A. AJCC Cancer Staging Manual. Sрringer, 7th Edition. 2011: 646.
7. Elder D.E. Lever "s Histopathology of the Skin. Wolters Kluver, 11th Edition. 2014: 1544.
8. Epstein J., Netto G. Biopsy Interpretation of the Prostate. Lippincott Williams and Wilkins, 5th Edition. 2014: 450.
9. Erickson L. Atlas of Endocrine Pathology. Sрringer, 1-st Edition. 2014: 178.
10. ESMO Minimum Clinical Recommendations for diagnosis, treatment and follow-up of cancers of unknown primary site. Annals of Oncology. 2005; 16 (S.1): 175-176.
11. Fletcher C.D.M., Bridge J.A., Hogendoorn P., Mertens F. WHO Classification of Tumors of Soft Tissue and Bone. WHO Press, 4th Edition. 2013; 5: 427.
12. Kurman R.J., Carcangiu M.L., Herrington C.S., Young R.H. WHO Classification of Tumors of the Female Reproductive Organs. WHO Press, 4th Edition. 2014; 4: 316.
13. Louis D.N., Ohgaki H., Wiestler O.D., Cavenee W.K. WHO Classification of Tumors of the Central Nervous System. WHO Press, 4th Edition. 2007; 1: 312.
14. Malpica A., Euscher E. D. Biopsy Interpretation of the Uterine Cervix and Corpus. Wolters Kluver, 2nd Edition. 2015: 368.
15. Nishizuka S., Chen S. T., Gwadry F. et al. Diagnostic markers that distinguish colon and ovarian adenocarcinomas: identification by genomic, proteomic, and tissue array profiling. Cancer Research. 2003; 63: 5243-5250.
16. Nordi Q.C. http://www.nordiqc.org.

There are many ways to diagnose cancer today. One of the most reliable methods for detecting oncological pathologies is the IHC study. What it is? Immunohistochemistry (abbreviated as IHC) allows high-precision detection of malignant cells in the material obtained using a biopsy. How is this test done? And how to decipher its results? We will consider these questions in the article.

Description of the procedure

Scientists have found that malignant tumor cells produce specific proteins - antigens. The immune system perceives these substances as foreign agents and begins to produce antibodies against them. Thus, the human body tries to fight the tumor.

The IHC study is based on the reaction of antigens and antibodies. What it is? During immunohistochemistry, a piece of tissue is taken for analysis. It is treated with antibodies to various types of malignant cells. Then the material is examined under a microscope and the reaction is observed.

If a piece of tissue contains cancer cells, then their antigens interact with antibodies. In this case, a glow (fluorescence) is noted in the biomaterial. This is a sign of the presence of malignant cells. This method allows you to detect cancer with 100% accuracy.

Analysis method

The patient is biopsied before the IHC study. What it is? Under local anesthesia, a piece of tissue is pinched off from the tumor (biopsy). This material is examined under a microscope. In some cases, tissue removed during a surgical or endoscopic operation is taken for analysis.

Then the biopsy is defatted and treated with paraffin. This is necessary for the safety of the material. The result is paraffin blocks, which are then cut into extremely thin layers.

The obtained sections are treated with preparations with antibodies to various types of malignant cells. The following assay kits can be used:

1. Small panel. This kit contains up to 5 antibodies.
2. Large panel. It may contain from 6 to several dozen types of antibodies.

The choice of the required antibodies for research depends on the intended diagnosis. In the direction of the analysis, the attending physician indicates which types of antigens must be identified in the patient. Depending on this, reagents for the test are selected.

You can get the results of the analysis on hands after 1 - 2 weeks after the study.

What the analysis shows

Often, patients are prescribed a histogram and an IHC study at the same time. What is it, and how do these two types of analyzes differ? With a histogram, the biomaterial is examined under a microscope and the presence of cancer cells is determined. In an immunohistochemical study, malignant neoplasms are detected using the antigen-antibody reaction. This method of research is considered more informative.

IHC shows not only the presence of atypical cells. With the help of this study, you can identify:

• type of neoplasm;
• the spread of the oncological process;
• location of the primary metastatic tumor;
• the stage of the cancer;
• tumor growth rate.

In addition, immunohistochemistry can be used to determine the sensitivity of cancer cells to chemotherapy drugs and radiation. This allows you to choose the most effective treatment method.

Indicators

In decoding IHC studies indicate the concentration of antigens of malignant neoplasms. Normally, such proteins should be completely absent. Their presence in biomaterial indicates the development of a tumor. The higher the concentration of antigens, the more active the growth of the neoplasm.

In addition, the analysis results indicate the type of atypical cells and their number. Only an oncologist can accurately interpret the test data.

Breast cancer detection

This test is prescribed for suspected breast cancer. In this case, the IHC study helps to identify not only the presence of malignant cells, but also the sensitivity of the neoplasm receptors to hormones. Indeed, often the growth of a breast tumor depends on the level of estrogen and progesterone. This study can also assess the need for hormonal medication.

A special set of markers is used to diagnose breast cancer. In decoding the analysis, it is necessary to pay attention to the concentration of the Ki-67 antigen. This indicator can be used to judge the prognosis of the disease, because it indicates the rate of tumor growth.

Antigen concentration is determined as a percentage. Depending on the Ki-67 level, a treatment method is selected:

1. Less than 15%. The prognosis of the disease is considered favorable. It can be cured with hormone therapy.
2. From 16 to 30%. Moderate growth of the neoplasm. The spread of the tumor can be halted with hormones.
3. Over 30 %. This concentration of antigen indicates the rapid growth of the cancer. Chemotherapy is required.

If the antigen indicator is less than 10%, then in the vast majority of cases, tumor growth can be stopped with the help of hormones. The concentration of Ki-67 over 90% is considered extremely dangerous. This indicates an advanced stage of cancer. In such cases, the disease is most often fatal.

Application in gynecology

Often, doctors prescribe an IHC study for patients with cervical dysplasia. This pathology is accompanied by the degeneration of healthy cells into atypical ones. Doctors assess this condition as precancerous. Grade 3 dysplasia almost always leads to the formation of a malignant tumor on the cervix.

IHC helps assess the risk of developing cancer in patients with dysplasia. For research, antibodies to p16INK4a and Ki-67 proteins are used.

The analysis determines not only the concentration of antigens, but also the ratio between the levels of the above proteins. The reference values \u200b\u200bof the research indicators are individual and depend on the alleged diagnosis. Only the attending physician can accurately decipher the test results. High levels of antigen concentration and the ratio between p16INK4a and Ki-67 indicate a high risk of developing cervical cancer.

Where can I take

Where is the IHC study carried out? Such a test can be done in many commercial medical centers and laboratories (for example, in "Invitro" or "Gemotest"), as well as in large oncological medical institutions. The analysis does not require the patient to stay in the hospital, except for those cases when the study is carried out after surgery.

Immunohistochemistry is not included in the list of services that are provided under the compulsory medical insurance policy. Therefore, such an analysis can only be submitted for a fee. Its price varies from 4,000 to 20,000 rubles. The cost of the test depends on the amount of antigens to be detected in the biopsy. The more markers indicated in the direction for analysis, the higher the cost of the study.

Alternative names: IHC, English: Immunohistochemistry or IHC.

Immunohistochemical study is a special method for diagnosing tumor diseases. The essence of the method is to study under a microscope tissue images previously treated with specific antibodies.

Currently, antibodies have been created for almost all tumors.

With the help of an immunohistochemical study, the following is performed:

Indications for immunohistochemistry

Any tissue can be examined using this method. The main indication for conducting is a suspicion of a tumor process.

The following indications for IHC were determined:

Contraindications

There are no contraindications to this research method. It is impossible to conduct it only if there is no way to obtain a biopsy material.

How is the immunohistochemical study performed?

The research itself consists of four stages:

The results of the study become known after 7 days (for a standard study - "small panel") or after 15 (extended study - "large panel").

Interpreting Results

The samples are examined by a pathologist with special training in IHC. In conclusion, the doctor notes to which antibodies the tissue tropism (affinity) is determined. Additionally, the morphological structure of the sample is described - which cells and how many are present.

Identification of tissue affinity for certain standard tumor antibodies indicates a specific type of cancer.

Additional Information

Immunohistochemical study is currently the most accurate method for diagnosing tumor diseases. It allows you to make a final diagnosis with an accuracy of 99%, determine the type of tumor, and identify its primary localization.

Literature:

Immunohistochemical study for breast cancer is an analysis that allows you to determine the sensitivity of a malignant neoplasm to certain substances. With its help, they diagnose the stage of pathology and find out if the treatment was prescribed correctly.

What is IHC?

If we talk about what is IHC oncology, then this is a study that allows you to identify the interaction between proteins and antigens. The fact is that cells affected by oncology synthesize protein. It has a connection with antibodies. With this in mind, experts use analysis to determine the shape and structure of the tumor.

Why is IHC carried out?

IHC is carried out in order to:

• determining the type of disease for the appointment of therapy;
• disclosure of the presence of metastases coming from the tumor, and their boundaries;
• detecting the source of metastases;
• diagnosing the stage of pathology when it is detected;
• evaluating the effectiveness of therapy;
• establishing the rate of development of metastases, if any;
• revealing the sensitivity of blood cells to certain drugs and methods of treatment in order to leave effective means and exclude those medicines and methods of therapy that do not affect the tumor.

How is the analysis done?

Immunohistochemistry of breast cancer is performed by sampling tissue affected by oncology. Biomaterial is obtained using a biopsy or during surgery. After that, the analysis is performed in the following stages:

• the biomaterial is placed in formalin, after which it is sent to the laboratory for further manipulations;
• specialists degrease the fabric;
• a special liquid paraffin is added to the material, which helps to reveal the structure of tumor cells;
• microtrauma is performed - the procedure for cutting off a thin layer of paraffin, 1 mm thick;
• the samples taken are placed on special glasses;
• the material is stained with reagents;
• the samples are left for 12 days, after which the result is evaluated.

Several types of markers are used for the analysis. If, during the study, an increased concentration of estrogens and progesterones is found in the biomaterial, a conclusion is made about the development of a tumor. This also indicates the appearance of metastases from a neoplasm. If the analysis showed average results, then they speak of weak tumor growth. This suggests that the patient, after appropriate therapy, has every chance of recovery.

Purely HER2 receptors affect the color gamut of reactors with cells that are affected by cancer. The more there are, the brighter the tone. In this case, histology manifests itself as an imprecise science, since the perception of shades of samples is subjective and depends only on the feelings of the specialist performing the analysis.

Decoding the results

When decoding the results, the receptors for progesterone - PR and estrogen - ER are taken into account. IHC in oncology also shows the content of the human epidermal growth factor receptor (HER2 protein) in the biomaterial taken as a sample for the study. Women with breast carcinoma have an elevated HER2.

Deciphering the results for estrogen and progesterone - if receptors for male and female hormones are found in the tumor, then the growth of education occurs under their influence, if PR and ER in the body are present in the norm or their concentration is increased. With timely qualified treatment, the patient can count on a favorable outcome.

Interpretation of the IHC results is performed depending on the shade of the samples on a special scale:

• 0-1 +. This result indicates that the protein concentration is not exceeded. Pathology therapy is not required.
• 2+. The value indicates the average protein concentration. This indicates the presence of a negative tumor (carcinoma) of the breast. In this case, it is recommended to perform another analysis, since the first study does not always give an accurate result. The FISH test is performed to clarify the diagnosis. This study determines the concentration of protein in each cell. With a normal content of HER2, the analysis will show a positive result, and with an increased content - negative.
• 3+. Protein exceeds normal value. A positive breast tumor is diagnosed.

If the analysis showed a positive tumor, then the pathology has an aggressive nature of development. It grows several times faster than with a negative value.

After studying the study, the result is decoded based on the indicator of the Ki67 marker. If its value is within the normal range, that is, 15-17%. When its level rises to 35%, the tumor begins to grow rapidly. Chemotherapy helps to slow down the process. If the analysis showed 85%, doctors refuse to treat the patient, since it is impossible to reduce the marker in this case and a fatal outcome cannot be avoided.

There are 4 types of cancer:

• luminal A - receptors for ER positive, negative for protein, Ki67 less than 14%;
• luminal B - receptors for ER, for HER2 - negative, Ki67 more than 15%;
• Erb B2 overexpressing - PR and ER - negative, protein - positive;
• Basal-like - negative value to progesterone and other factors.

Prognostic factors

Such factors demonstrate the possible behavior of the tumor at the time of analysis. Treatment has no effect on the study. Thanks to IHC, the doctor sees how the tumor behaves. The specialist uses the prognosis to determine the appropriate therapy for each patient.

Predictive factors

Breast cancer IHC studies can determine the effectiveness of treatment. If the test shows a positive cancer, then chemotherapy is prescribed. If the value is negative, medication is taken.

IHC is a study that allows you to determine the type of breast cancer and prescribe adequate treatment, which can save the patient's life, and in some cases, get rid of the pathology.