Detection of Calreticulin as a Candidate Prognostic Biomarker  in Invasive Breast Carcinoma from a Biological Scaffold-Based  3D Co-culture System
	    		
	    			
	    			
		        		
			        		
		        		
			        
		   		
		   		
		   			
		   		
	    	
    	 
    	https://doi.org/10.47836/mjmhs.19.1.24
   		
        
        	
        	
        	
        		- Author:
	        		
		        		
		        		
			        		Mohammad Mehdi Sabaghpour Azarian
			        		
			        		
			        		
			        			1
			        			
			        		
			        		
			        		
			        		
			        		;
		        		
		        		
		        		
			        		Norazalina Saad
			        		
			        		
			        		
			        			2
			        			
			        		
			        		
			        		
			        		
			        		;
		        		
		        		
		        		
			        		Aslah Mohamad
			        		
			        		
			        		
			        			3
			        			
			        		
			        		
			        		
			        		
			        		;
		        		
		        		
		        		
			        		Rozita Rosli
			        		
			        		
			        		
			        			2
			        			
			        		
			        		
			        		
			        		
			        		
		        		
		        		
		        		
		        		
		        			
			        		
			        		Author Information
			        		
		        		
		        		
			        		
			        		
			        			1. Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang,  Selangor, Malaysia
			        		
			        			2. Selangor, Malaysia. 2  UPM-MAKNA Cancer Research Laboratory, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor,  Malaysia
			        		
			        			3. Aquatic Animal Health and Therapeutics Laboratory, Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang,  Selangor, Malaysia
			        		
		        		
	        		
        		 
        	
        	
        	
        		- Publication Type:Journal Article
 
        	
        	
        		- Keywords:
        			
	        			
	        				
	        				
			        		
				        		Calreticulin, Breast cancer, Prognostic marker, 3D co-culture system
			        		
			        		
	        			
        			
        		
 
        	
            
            
            	- From:Malaysian Journal of Medicine and Health Sciences
	            		
	            		 2023;19(No.1):173-180
	            	
            	
 
            
            
            	- CountryMalaysia
 
            
            
            	- Language:English
 
            
            
            	- 
		        	Abstract:
			       	
			       		
				        
				        	Introduction: Breast cancer is the most common cancer in women and the world’s second leading cause of death 
in women, after lung cancer. Calreticulin (CRT), an endoplasmic reticulum (ER) multipurpose protein, has been 
proposed as a potential biomarker for breast cancer. However, reports on the correlation between CRT expression 
and cell invasiveness in breast cancer micro-tissues are scarce. Thus, in the current study, we analyzed the potential 
correlation between CRT and invasiveness of breast cancer in a biological scaffold-based 3D co-culture system. 
Methods: MCF7, MDA-MB-231 and MCF-10A breast cell lines were co-cultured in a 3-dimensional (3D) system with 
MRC-5 lung fibroblast cell line in the cell density ratio of 3:1. Thereafter, calreticulin gene and protein expression 
levels were determined based on quantitative reverse transcription-polymerase chain reaction (RT-qPCR) and immunohistochemistry, respectively. Moreover, via RT-qPCR analysis, the gene expression levels of calreticulin-related 
candidate metastasis genes in breast cancer micro-tissues were carried out. Results: The results showed occasional 
foci of lumen-like morphology in the non-cancerous breast micro-tissues and the formation of solid clusters for 
breast cancer micro-tissues. Moreover, immunohistochemistry results revealed protein expression of calreticulin in 
non-cancerous and cancerous breast micro-tissues with cytoplasmic and nucleic acid localizations. It was found that 
PCMT1 and ER-α genes were significantly downregulated (p < 0.01) in invasive breast cancer micro-tissues. Conclusion: This study suggests that CRT and CRT-related candidate metastasis genes may potentially serve as prognostic 
biomarkers in invasive breast carcinoma. 
				        	
				        
				    
			    
 
	        
	        
	        	- Full text:11.2023my1424.pdf