RU2466390C2 - Method for prediction of developing endometrial cancer in pathological endometrial processes in females of reproductive age - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to a method for prediction of developing endometrial cancer consisting in detecting clinical signs and methylation of MLH1, RASSF1, GSTP1, p16, RAR-b, CDX1 genes, and calculating a probability factor of developing endometrial cancer. If the value p is more than 0.6, a high probability of developing endometrial cancer is predicted; the value p equal to 0.3-0.59 show a moderate probability of developing endometrial cancer, while the value p less than 0.29 provides a low probability of developing cancer.

EFFECT: method provides accuracy of prediction of developing endometrial cancer in pathological endometrial processes in females of reproductive age.

2 ex

Description

The invention relates to medicine, in particular to gynecology. One of the ways to predict cancer of the uterine body is to identify and evaluate the following risk factors: impaired carbohydrate metabolism, arterial hypertension, recurrent endometrial hyperplastic processes, polycystic ovary syndrome, infertility, change in ultrasound parameters (M-echo, blood flow velocity) (Chulkova O.V. Precancer and endometrial cancer in women of reproductive age: Diss. ... Doctor of Medical Sciences. - M., 2003. - 268 p.).

The disadvantage of this method is the low accuracy due to the identification, for the most part, of general clinical risk factors that are characteristic not only for malignancy of pathological processes of the endometrium.

Closest to the proposed method is predicting the development of cancer of the uterus in pathological processes of the endometrium based on the identification of methylation of the hMLH1 and APC genes in the endometrium (Shalkova M.Yu. Endometrial cancer: methylation of the hMLH1 and APC genes as risk factors and progression pattern: the Dissertation ... of the candidate of medical Sciences - Kiev, 2008 - 141 p.).

The author notes an increase in the frequency of epigenomic disorders in the series endometrial hyperplasia → recurrent endometrial hyperplasia → atypical endometrial hyperplasia → endometrial cancer.

The disadvantage of this forecasting method is its low sensitivity and specificity, probably due to the determination of methylation of only two tumor suppressor genes, without a comprehensive assessment of clinical risk factors.

The objective of the invention is a method for improving the accuracy of predicting the development of cancer of the uterus.

The problem is solved by a method for predicting the development of cancer of the uterus in pathological endometrial processes in women of reproductive age, including the determination of gene methylation in an endometrial biopsy, characterized in that it determines the clinical signs and methylation of the MLH1, RASSF1, GSTP1, p16, RAR-b, CDX1 genes, calculate the coefficient of probability of developing cancer of the uterus according to a formula developed on the basis of binary logistic regression:

where p is the coefficient of probability of developing cancer of the uterus (from 0 to 1);

e is the base of the natural logarithm and is equal to 2.7182818282845904,

z = -11.2754 + 2.32327; X1 + 2.713726; X2 + 2.631425; X3 + 2.412264; X4 + 2.571489; X5 + 2.411352; X6 + 2.521; X7-3.521352 X8 + 1.132435 X9 + 0.85648 X10 + 1.32356 X11 + 0.765356 X12 + 1.211383 X13 + 1.234 X14 + 0.921237 X15 + 0.516623 X16,

X1 - morphotype (value: endometritis - 1, simple hyperplasia without atypia - 2, complex hyperplasia without atypia - 3, endometrial polyp - 4, complex hyperplasia with atypia - 5).

X2 - methylation RASSF1 (value: in the presence of 1, in the absence of 0)

X3 - methylation of MLH1 (value: in the presence of 1, in the absence of 0)

X4 - p16 methylation (value: in the presence of 1, in the absence of 0)

X5 - RAR-b methylation (value: in the presence of 1, in the absence of 0)

X6 - methylation GSTP1 (value: in the presence of 1, in the absence of 0)

X7 - methylation Cdx1 (value: in the presence of 1, in the absence of 0)

X8 - index of resistance to blood flow in the arcuate arteries (value 0.3-0.65)

X9 - violation of carbohydrate metabolism (value: in the presence of 1, in the absence of 0)

X10 - hypertension (value: if present - 1, if not - 0)

X11 - recurrence of endometrial hyperplastic processes (value: in the presence of 1, in the absence of 0)

X12 - uterine fibroids and / or adenomyosis (value: in the presence - 1, in the absence - 0)

X13 - hereditary burden of tumor diseases (value: in the presence of 1, in the absence of 0)

X14 - polycystic ovary syndrome (value: in the presence of 1, in the absence of 0)

X15 - infertility (value: if present - 1, if not - 0)

X16 - degree of obesity (value: 1-3)

and with a value of p greater than 0.6, a high probability of developing cancer of the uterine body is predicted, with p equal to 0.3-0.59, a moderate probability of developing cancer, with p below 0.29, a low probability of developing cancer.

The sensitivity of the method is 98%, specificity is 92%. The coefficients were calculated on the basis of a comprehensive examination and treatment of 216 patients with pathological endometrial processes. After processing and analysis of the obtained data (52 evaluated parameters) in the Microsoft Excel programs, and the Statistica for Windows application package v.7.0, StatSoft Inc. (USA), statistically significant risk factors were determined and their contribution (coefficient in the formula of binary logistic regression) to the probability of developing cancer of the uterus (N.I. Pirogov. Logistic regression. Basic concepts and capabilities of the Method. Http: //www.biometrica tomsk.ru/logit_1.htm).

In practice, the method is as follows.

When examining a patient (on an outpatient basis or in a hospital), the presence of clinical and anamnestic risk factors for developing cancer of the uterine body is determined: impaired carbohydrate metabolism or diabetes mellitus, arterial hypertension, recurrence of endometrial hyperplastic processes (GPE), myoma and / or adenomyosis, hereditary burden of tumor diseases , polycystic ovary syndrome (PCOS), infertility, obesity, ultrasound with CDK.

After curettage of the uterus, part of the material is sent for morphological examination, and part for DNA testing.

Methylation of tumor suppressor genes was determined by the method of methyl-sensitive polymerase chain reaction (MCH-PCR).

For the convenience of everyday use, the formula is entered once in Microsoft Office Excel. Next, data is entered into the cells. As a result, the risk of malignant transformation of the endometrium for a particular patient was calculated.

An example program is presented below.

Based on the data obtained, 3 groups of patients can be identified:

- with a low probability of developing cancer - p = 0.0-0.29

- with a moderate probability of developing cancer - p = 0.3-0.59

- with a high probability of developing cancer - p = 0.6-1.0

The sensitivity of the method is 98%, specificity is 92%.

Example 1

Patient S. (No. 17), 39 years old, was hospitalized with suspected ultrasound process of the endometrium. The examination revealed clinical risk factors: arterial hypertension, recurrence of HPE, combined uterine pathology (myoma and adenomyosis), hereditary burden of tumor diseases. Histological examination revealed atypical endometrial hyperplasia. Methylation of genes RASSF1, GSTP1, p16 was detected. IR with ultrasound = 0.55.

Thus: X = 5, X2 = 1, X3 = 0, X4 = 1, X5 = 0, X6 = 1, X7 = 0, X8 = 0.55, X9 = 0, X10 = 1, X11 = 1, X12 = 1, X13 = 1, X14 = 0, X15 = 0, X16 = 0.

After entering the data into the formula, a high probability of developing cancer of the uterine body was obtained (p = 0.82), which allowed the patient to be assigned to a high risk group for developing cancer of the uterus.

Treatment was proposed according to the scheme of initial forms of cancer, which the patient refused. Conducted standard treatment with progestogens according to the standard scheme.

After 6 months, the patient was delivered for emergency reasons with a diagnosis of "Uterine bleeding." Produced curettage of the uterus. Diagnosed with endometrial cancer.

Example 2

Patient M. (No. 94), 43 years old, was hospitalized in a hospital with complaints of spotting from the genital tract. An examination revealed risk factors: hereditary burden of tumor diseases, infertility. Hysteroscopy, polypectomy, curettage of the uterus were performed. Diagnosed with fibrous-glandular endometrial polyp. No methylated tumor suppressor genes were detected. IR with ultrasound = 0.49.

Thus: X1 = 4, X2 = 0, X3 = 0, X4 = 0, X5 = 0, X6 = 0, X7 = 0, X8 = 0.49, X9 = 0, X10 = 0, X11 = 0, X12 = 0, X13 = 1, X14 = 0, X15 + 1, X16 = 0.

After the data were entered into the formula, a value of p = 0.26 was obtained, which allowed the patient to be assigned to the low risk group for developing uterine cancer. The patient is recommended for dynamic observation. In the control study (ultrasound) after 6 and 12 months of relapse was not noted.

Example 3

Patient V. (No. 2), 34 years old, was hospitalized with a diagnosis of "Recurrent endometrial hyperplasia." A history of 2 curettages for uterine bleeding. Histologically, simple endometrial hyperplasia was detected. Progestogen treatment was performed.

Clinical risk factors were identified: arterial hypertension, recurrence of HPE, hereditary burden of tumor diseases, obesity 2 tbsp.

Produced curettage of the uterus. Morphologically revealed complex endometrial hyperplasia without atypia. PCR detected methylation of the RAR-b and CDX1 genes. IR with ultrasound = 0.48.

Thus: X1 = 3, X2 = 0, X3 = 0, X4 = 0, X5 = 1, X6 = 0, X7 = 1, X8 = 0.48, X9 = 0, X10 = 1, X11 = 1, X12 = 0, X13 = 1, X14 = 0, X15 = 0, X16 = 2.

After entering the data into the formula, the result was p = 0.78, which allowed the patient to be assigned to the high-risk group.

The treatment was carried out according to the scheme of complex hyperplasia with atypia (GnRH agonists) - 6 months.

Produced a control curettage - no pathology was detected.

Claims (1)

A method for predicting the risk of developing cancer of the uterus in pathological processes of the endometrium in women of reproductive age, including determining methylation of genes in an endometrial biopsy, characterized in that it determines the clinical signs and methylation of the MLH1, RASSF1, GSTP1, p16, RAR-b, CDX1 genes, and calculate the coefficient the probability of developing cancer of the uterus according to the formula:

where p is the coefficient of probability of developing cancer of the uterus;
e - the base of the natural logarithm and equal to 2.7182818282845904,
z = -11.2754 + 2.32327 * X1 + 2.713726 * X2 + 2.631425 * X3 + 2.412264 * X4 + 2.571489 * X5 + 2.411352 * X6 + 2.521 * X7-3.521352 * X8 + 1.132435 * X9 + 0.85648 * X10 + 1.32356 * X11 + 0.765356 * X12 + 1.211383 * X13 + 1.234 * X14 + 0.921237 * X15 + 0.516623 * X16,
X1 - morphotype (value: endometritis - 1, simple hyperplasia without atypia - 2, complex hyperplasia without atypia - 3, endometrial polyp - 4, complex hyperplasia with atypia - 5)
X2 - methylation RASSF1
X3 - methylation of MLH1
X4 - p16 methylation
X5 - RAR-b methylation
X6 - methylation GSTP1
X7 - methylation CdX1
X8 - index of resistance to blood flow in the arcuate arteries
X9 - violation of carbohydrate metabolism
X10 - hypertension
X11 - recurrence of endometrial hyperplastic processes
X12 - uterine fibroids and / or adenomyosis
X13 - hereditary burden of tumor diseases
X14 - Polycystic Ovary Syndrome
X15 - Infertility
X16 - the degree of obesity (1, 2, 3),
and with a value of p greater than 0.6, a high probability of developing cancer of the uterine body is predicted, with p equal to 0.3-0.59, a moderate probability of developing cancer, with p below 0.29, a low probability of developing cancer.