




i hc Khoa hc T nhiên
 ngành: ; 60 42 30
PGS. TS. 
 2011

Abstract: 
              

 

    

-


Keywords: ; Gen CXCL12; ; 

Content
1.Mở đầu


 -80%.
  

y 


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




Hóa chất
-                

- Enzyme: Proteinase K, Fastdigest® Msp
- -f, U-r, M-f, M-
- 
- 
- 
- Enzyme CpG Methyltrasferase (M.SssI) (N
- 
- 
- dNTPs 
- Phenol, Chloroform, Isopropan
- 
3
, Na
2
CO
3

- Acid boric, Tris-base, ETDA, ethanol,
- 

Msp 
-
o
C.
2.5. Khuếch đại vùng promoter của gen CXCL12 bằng kỹ thuật MSP
Xử lý bisulfite ADN bằng kit DNA Methylation-Gold TM kit và xử lý bisulfite ADN
theo phương pháp của Herman va
̀
cs (1996) [2]
Thực hiện phản ứng PCR để khuếch đại gen CXCL12

                
Wendt và cs, (2006) [3]            
MethPrimer.

CXCL12 
CXCL12-
- f ): - GGA GTT TGA GAA GGT TAA AGG TC - 
- r) :- TTA ACG AAA AAT AAA AAT AGA CGA T - 
CXCL12-
-- GAG TTT GAG AAG GTT AAA GGT TGG - 
- - TAA CAA AAA ATA AA ATA CAA CAA T - j
2.6. Phân tích kết quả bằng phương pháp thống kê
2

 CXCL12 và tình

tràng.
3. Kết quả.


Hình3. ảnh điện di sản phẩm cắt ADN bằng enzyme MspI với 3 băng 302bp, 202bp và
100bp
(Điện di trên gel polyacrylamide 10% và nhuộm bạc)

enzyme MspI.
 enzyme MspI 


. 

                   
enzyme MspI 
N

n
3.1.4. Phân tích số liệu bằng phương pháp thống kê
Q


CXCL12 trong ung

3.2. Kết quả phân tích MSP


 6

Hình4. ảnh điện di ADN tổng số tách từ mô

Hình 6. Kết quả khảo sát đảo CpG sử dụng phần mềm cpgplot (EMBOSS)[4]
CPGPLOT islands of unusual CG composition
3001-6001 from 1 to 3001
Observed/Expected ratio > 0.60
Percent C + Percent G > 50.00
Length > 100
Length 297 (137 433)
Length 161 (623 7833)
Length 225 (891 1115)
Length 1346 (1174 2519)
Length 341 (2526 2866)
Kết quả khuyếch đại đoạn trình tự nằm trong đảo CpG thuộc vùng promoter của
gen CXCL12 bằng kỹ thuật MSP

8

Hình7. ảnh điê
̣
n di sản phẩm PCR cu
̉
a gen CXCL12
(điê
̣
n di trên gel polyacrylamide 10% và nhuộm bạc)
-
- 
1,3,5,7: UMethyl.

                
promoter cCXCL12.

U
M9
Hình 8.Tỷ lệ methyl và unmethyl khảo sát trên 2 loại mô u và lân cận u
U: unmethyl; M: methyl
 (hình 13) 
,  
(43,75
(62,5%).

CXCL12


, 
 có 
  
methyl hóa hoàn toàn

có 
Kết Luận

-CXCL12:

CXCL12 

(p>0,05).
2. 








CXCL12  

CXCL12 


References
Tài liệu tiếng Việt
1. , Di truyền ung thư 
tr.12.
2,  hóa
a gen CXCL12  Tạp chí y học Việt
Nam (2), tr. 72-77.
3. ), Cơ sở di truyền học phân tử và tế bào, Nhà

4 
- 
MLH1 y
Tạp chí khoa học   27(2), tr 315.
Tài liệu Tiếng Anh

10
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