Breast cancer patients under 45 years of age using tissue microarray technique Role of immunohistochemical biomarkers

Breast cancer patients under 45 years of age using tissue microarray technique

Role of immunohistochemical biomarkers

Purpose: Tissue microarray (TMA) is used for the paraffin block of tumor tissue in patients under 45 years of age.
Expression status of five biomarkers such as estrogen receptor (ER), progesterone receptor (PR), HER-2, COX-2 and Survivin
And to investigate the significance as a prognostic factor in patients.
Materials and Methods: From March 1994 to August 2005, TMA analysis was performed among breast cancer patients under 45 years of age
A total of 212 patients were enrolled. The age of the patients ranged from 23 to 45 years and the median age was 39 years.
The probing period was 24 months and the median follow - up period was 60 months. After selecting a patient under 45 years of age,
After marking the cancerous area through the dyed slides, a microstructure array was made through a TMA instrument. And five living bodies
Immunohistochemical staining of the markers was performed and the pathologist read it. The results and the age,
The results of this study were as follows: 1) Patients were followed up for 2 months.
The post - factors were analyzed.
Results: Patients were divided into two groups according to T stage: T1 (90 patients), T2 patients (101 patients), 105 patients (49.5%), axillary lymph nodes
. The 5-year disease-free survival and 5-year survival rates of all patients were 82.3% and 90.4%, respectively.
Recurrence or distant metastasis occurred during follow - up and 20 patients died of cancer. ER, PR, HER-2, COX-2,
The rate of positive expression of survivin was 38.2%, 45.3%, 25.9%, 41.5%, and 43.4%, respectively. The tumor size,
The presence of metastasis, hormone receptor status, and HER-2 status are important prognostic factors for univariate analysis of disease-free survival
And the effect on survival rate was tumor size, lymph node metastasis, hormone receptor, and HER-2 expression.
COX-2 and survivin did not play a role as prognostic factors, but HER-2 and COX-
Bad tendency could be observed. Multivariate analysis showed that only T-stage and lymph node metastasis were important prognostic factors and ER
Had the meaning of the boundary.
CONCLUSION: Hormone receptor status and HER-2 were prognostic factors in women under 45 years of age and COX-2 and survivin
I could not find a role as a later argument.
               

                                                        Object and method

1. Research subjects

 From March 1994 to August 2005 at Soonchunhyang University Hospital
Among the primary breast cancer patients under 45 years of age,
Of the paraffinic tissues were left. Para
After finely sectioning the pin block, H-E staining showed that the tumor was present.
Two cores were marked in the paraffin block of 214 patients.
It was commissioned by Isu ABGS Co., Ltd. Inspection Center
Using a machine, tissues with a diameter of 2 mm were collected and one
In the paraffin block, 48 tissues from 24 patients
I made TMA by soldering. After making TMA, re-H-E salt
A color sample was prepared and the presence of the tumor was confirmed.
No tumors were found in either of the two cores,
The tissues of both metachronous breast cancers were identified and excluded
Respectively. Ten had tumors on one side of the core,
You can get information from the core,
I could sit. The patient's age is between 22 and 45 years old.
The median was 39 years old and the age of 35 was under 63, accounting for 29.7%
Respectively. The surgical methods were modified radical mastectomy
And 19 patients underwent simple mastectomy or skin preservation
Mastectomy was performed. Breast conserving surgery was performed in 78 patients
The other two patients were treated with breast conserving surgery,
Cancer was performed by a modified radical mastectomy. Size of tumor
The postoperative pathologic findings were between 0.9 and 10 cm
T (DCIS) was classified into 6 patients (2.8%), T1 was 90
(42.5%), T2 101 (47.6%), and T3 15 (7.1%), with T1 and T2
(Table 1). Pathologic examination revealed bilateral
All 10 patients had breast cancer and all had invasive ductal carcinoma
(infiltrating ductal carcinoma). Tissue of unilateral breast cancer
The type of ductal carcinoma in situ (DCIS)
(Medullary carcinoma), tubular carcinoma
1 tubular carcinoma, 2 lobular carcinomas,
Paget disease in one patient and all remaining invasive tubular carcinoma
.
 All patients underwent axillary lymph node dissection.
There were 105 patients (49.5%) with a mediastinal metastasis and lymph node metastasis
The number of samples was ranged from 1 to 45 in a wide range. lymph
If there is one metastasis among 105 patients with metastasis
24 (23.1%), followed by three (21.2%), two
(12.5 percent), and nine (9.7 percent). Four lymph node metastases
In the high-risk group, 47 patients were treated with total lymph node metastasis
Of the total.
 All patients who underwent breast conserving surgery had postoperative radiation levels
Radiation therapy was performed by 4-MV
Type accelerator, and since 2000, 6-MV linear accelerator
Were used. 1440 after treatment of 5040 cGy / 28 fx in the former breast
cGy / 7 fx boost treatment and axillary lymph node metastasis
, 5040 cGy radiotherapy was applied to the lymph node area
Respectively. When mastectomy was performed, there were 4 lymph node metastases
Or when the size of the primary lesion is 5 cm or more, the chest wall and limb
5040 cGy treatment was performed at the metastatic site of the cervix. Anticancer drug
Is an anticancer therapy commonly used at the time of treatment
(regimen), the most commonly used anticancer drugs
cyclophosphamide, methotrexate and fluorouracil (CMF)

2. Method

 1) Fabrication of tissue microarray (TMA)

 Among the stored formalin-fixed paraffin-embedded breast cancer tissues
Re-search for H-E staining slides by searching for subjects aged 45 years or younger
And a pathologist to find a tumor site and place TMA in group 2
Respectively. Using the microarray array,
Pull it out into a 2 mm diameter cylinder and insert a new paraffin-
I moved to the rock. The TMA block is microtome-shaped with a thickness of 3-5 μm
After cutting, spread the wrinkles and attach to the slide in a certain direction
Lt; / RTI & gt; This process was carried out by Isu Abxis Co., Ltd.
 The fabricated microarray paraffin block was continuously
And immunohistochemical staining was performed.
 2) Immunohistochemical staining.
 (1) estrogen, progesterone receptor (ER, PR)
We observed more than 4 cases in the field of view,
According to the ratio of cells stained with color, when it is 10% or less, 0 point,
1 point for 11-25%, 2 points for 26-50%, 3 points for 51-75%
When it was 76% or more, it was divided into 4 points. Also, depending on the staining intensity
0 for voice, 1 for weak, 2 for moderate, 3 for strong
Respectively. The proportion of positive cells and nuclear staining intensity score
If the sum is 0, negative, 1 to 3 points are positive, 4 to 5 points are moderate
Positive, 6 to 7 were strongly positive, and moderate
Were classified as hormone receptor positive group.
 (2) c-erb B2 (HER-2): The readout indicates the cell membrane of tumor cells
Depending on the degree of staining, the score is 0, weak in some cells
1 point when showing staining, some cells but strong staining.
In most cases the cells are moderately stained at 2 points,
When the stain is strong, it is calculated as 3 points.

3) COX-2: The reading is the cell-positive staining of cells
0 point for 0%, 1 point for less than 10%, 2 points for 10 to 50%
3 points at 50 to 80% and 4 points at 80% or more, and the staining intensity
Negative 0 point, weak positive 1 point, moderate positive 2 point, strong positive 3 point
And the percentage of tumor cells stained positive
When the sum of color intensity scores is 0 to 6, COX-2 is negative,
And were read as COX-2 positive.
 (4) Survivin: The reading is the nucleus or cytoplasmic salt of tumor cells
And the ratio of colored cases was calculated. There are no stained areas or sheep
When the stained area was less than 5%, it was interpreted as negative
All. If more than 5% were stained, 1 point less than 30%, 30 ~
60% moderate 2 points, 60% or more 3 points, and statistics
Three points were classified as positive for treatment.
3. Statistical processing
 Clinical and pathologic characteristics of all patients, immunohistochemical staining
Results of follow-up and follow-up of patients after treatment
SPSS (Statistical Package for the Social Science) version 10
And analyzed. Correlation analysis is based on Pearson's Correlation
Coefficients were used and survival analysis was performed using the Kaplan-Meier method
The log-rank test was used to compare survival among the groups.
Respectively. For multivariate analysis, Cox Proportional Hazard
Model was used and when the p-value was less than 0.05,
The p value was between 0.05 and 0.1.
And that the meaning of the boundary is meaningful.
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