الفهرس 
Title pagOnly 14 pages are availabe for public view
 |  | from | 201 |  |  |
| | | from | 201 | | |
Abstract
Breast cancer is the most prevalent form of cancer in women all over the globe, and is one of the main causes of morbidity and mortality.
The complexity and heterogeneity of breast cancer demonstrated by gene expression profiling make its treatment challenging
Despite recent advances in therapeutic modalities which allowed better management of breast cancer, yet, the high recurrence rate and the potential to progress mark breast carcinoma as a health problem.
BCSCs are a tiny subset of cells capable of self-renewal and propagation of breast cancer cell populations that are diverse. The BCSCs hypothesis showed breast cancer’s cellular genesis, tumour maintenance, and progression.
The idea that stem cells in hypoxic environments depend on HIF activity to retain their undifferentiated phenotype is gaining traction. It’s worth noting that the existence of hypoxic regions inside a tumour may indicate potential habitats for cancer stem cells. It’s been suggested that the link between tumour hypoxia and poor patient outcomes could be due to an increase in the presence of cancer stem cells.The present study aimed at detecting ALDH1 and CAIX immunostaining in invasive ductal carcinomas and DCIS, if present, using immunohistochemistry and correlating them with clinicopathological parameters.
To achieve this aim, Formalin-fixed paraffin-embedded (FFPE) tumor tissue blocks of 75 patients who underwent who underwent mastectomy or wide local excision for invasive ductal carcinoma, in the period between January 2020 and May 2021. The speciemens were retrieved retrospectively from the archives of the Pathology department. The sections were cut and stained with ALDH1, and CAIX antibodies.
All patients received had available clinical data, as well as available ER, PR, and HER2 immunoreactivity results from the files of the pathology department.
Biopsies of recurrences of invasive ductal carcinoma, treated prior to tumor resection and with unavailable clinical data and/ or ER, PR, and HER2 immunoreactivity data, were excluded.
Measuring the proportion (P) of tumour cells exhibiting distinctive staining (from undetectable to homogenous staining, or 100 percent) and estimating the intensity (I) of staining were used to assess the results. Scoring of ALDH1 and CAIX immunostains, reactivities were assessed in terms of Quick score.
In the present study, the age of patients ranged between 26 and 84 years with a mean age of 54.16 years ± Std. deviation of 12.71 years. Forty nine (65.3%) cases were older than 50 years while, twenty six were younger than 50 years (34.7 %). Fifty seven percent of tumors were larger than 20 mm, whereas forty two percent were ranging in size from 0 to 20 mm. Most tumors (72%) were unifocal, and the remaining 28% were multifocal.
Out of the 75 studied cases, , 53 cases (70.67%) were classified as (G2) and 22 cases (29.33%) were classified as high grade tumors (G3).
Forty three cases (57.33%) revealed the presence of lymphovascular invasion. Regarding the changes in the adjacent non neoplastic breast tissue, out of the 75 cases, forty one cases (54.67%) revealed fibrocystic changes, atypical hyperplasia was detected in only (11) cases (14.67%) out of 75 studied cases.
DCIS was detected in 16 cases (21.33%) out of the 75 studied cases and necrosis was in 34.67% of the cases.
The cases were stratified into AlDH1 positive and ALDH1 negative groups according to the Quick score. Twenty nine cases (38.67%) were negative for AlDH1 immunostaining, while, 61.33% of cases (n=46) revealed positive AlDH1 immunostaining. The stromal cells also stained positive in some cases, as ALDH1 positive cytoplasmic staining was noted in stromal cells in 21 cases (28%). In these cases the percentage of positive cells ranged from 10% to 100 %.
Significant correlations were identified between ALDH1 protein expression and high histological, grade 3; (p=0.0001)] , and CAIX imunostaining (p=0.01), but not with other variables, including age, tumor size and focality, associated atypical hyperplasia, ER, PR, and HER2 immunoreactivity, lymphovascular invasion and necrosis.
Associated fibrocystic changes was noted in 41 cases out of the 75 studied cases, (72.41% ) of those 41 cases were ALDH1 negative and exhibited a significant inverse correlation with ALDH1 + immunostaining, being more common with ALDH negative invasive ductal tumors. (p=0.014)
Regarding CAIX immunostaining cases were also stratified into CAIX positive and CAIX negative groups according to Quick score. Forty two cases (56.00%) revealed CAIX negative immunostaining, while, (44.00%) of cases (n=33) revealed CAIX positive immunostaining. CAIX did not stain the tumor stroma in any of the studied 75 cases.
The CAIX positive immunostaining within tumor cells correlated significantly with tumor grade 3;( p=0.00124).
Triple-negative tumors were more likely to be CAIX+ than non–triple-negative tumors (72.73% versus 39.06%, P =0.03752). Conversely, ALDH1+ tumors had no significant differences observed in TNBC tumors.
Twenty-five cases (75.76%) out of 33 CAIX positive invasive ductal tumors compared with 21 cases (50.00%) out of 42 CAIX negative invasive cancers, had ALDH1+ tumor cells.
CAIX + tumors were noted in 8 cases out of 29 (27.59 %) within the ALDH1 negative tumors, and in 25 cases out of 46 (54.35%) within ALDH1 positive tumors.
There was a significant correlation between the overall ALDH1+ and CAIX+ immunostaining (Z=2.2737, p=0.0232).
In the current study, 16 (21.33%) of the 75 invasive cancers studied had associated DCIS. Within this subgroup of tumors, 7 cases (43.75%) of 16 had ALDH1+ cells and 5 cases (31.25%) of 16 had CAIX+ cells detected in foci of DCIS.
No significant difference was detected between tumors with DCIS and those cases without DCIS for both ALDH1 and CAIX immunostaining,
As for CAIX immunostaining, 7 cases (63.64%) of 11 CAIX negative invasive cancers with DCIS was ALDH1 negative, compared with 4 cases (36.36%) CAIX negative tumor cells showed ALDH1+ tumor cells detected in foci of associated DCIS.
Three cases (42.86) of 7 ALDH1 + invasive cancers with DCIS showed positive immunoreactivity for CAIX immunostaining, while 4 cases (57.14%) out of 7 was CAIX negative.
The significant correlation between ALDH1 immunostaining and CAIX immunostaing was lost within the subgroup tumors with DCIS.
No significant difference was detected for ER, PR, HER2 or TNBC status with ALDH1+ immunostaining. Conversely, CAIX+ staining was more common in TNBC (P =0.03752), but did not show a significant difference with ER+, PR+ OR HER2+ status
In conclusion, our findings revealed that the ALDH1 breast CSC marker is localised according to histologic grade and intratumoral microenvironment. Our investigation revealed the presence of all ALDH1+ stem cell markers in DCIS linked with invasive malignancy, confirming earlier research that indicated tumour stem cell markers were related with clinicopathologic criteria of tumour aggressiveness such as high tumour grade.
Furthermore, ALDH1+ tumour cells were shown to be more prevalent in areas of hypoxia in this research, while ALDH1+ tumour cells were found to be more often linked with areas of high microvascular density in previous investigations.
This research backs up the idea that the tumour microenvironment influences breast cancer stemlike cells’ phenotypic plasticity, and it has significant clinical implications for the development of treatment methods targeting cancer cells with stem-like characteristics.