c-erbB-2反义寡脱氧核苷酸联合EGF对乳腺癌细胞的影响

Effects of c-erbB-2 antisense oligodeoxynucleotides combine EGF for breast cancer

目的:探讨c-erbB-2反义寡脱氧核苷酸(antisense oligodeoxynucleotides,ASODN)联合表皮生长因子(epidermal growth factor,EGF)对乳腺癌SK-Br-3细胞的影响。方法:c-erbB-2反义、正义和无义寡脱氧核苷酸用99mTc标记,联接EGF形成99mTc-ODN-EGF,测定乳腺癌细胞SK-Br-3的摄取率和返流比率。将99mTc-ODN-EGF加入细胞培养,5×104cpm/d,给药3 d,检测细胞活力和c-erbB-2蛋白表达。结果:240 min时99mTc-ASODN-EGF9、9mTc-S-ODN-EGF9、9mTc-NODN-EGF和99mTc-ASO-DN的细胞摄取率分别为23.70%、25.24%、24.91%和5.30%,EGF三组药的摄取率明显高于99mTc-ASODN,P=0.0629。9mTc-ASODN-E-GF9、9mTc-SODN-EGF和99mTc-NODN-EGF12 h的细胞返流比率分别为51.28%、75.70%和81.20%,反义寡核苷酸组明显降低,P=0.002。以上四组药物作用细胞后,细胞活力分别为0.51±0.13、2.33±0.48、2.57±0.57和2.12±0.45,反义寡核苷酸组的细胞活力明显低于其余三组,P值分别为0.009、0.014和0.036;c-erbB-2蛋白表达分别为118.77±5.63、134.16±3.88、134.76±1.76和131.24±6.66,99mTc-ASODN-EGF组蛋白表达明显降低,P=0.013。结论:99mTc-ASODN-EGF的细胞摄取率高,返流比率低,使乳腺癌细胞活力降低,c-erbB-2蛋白表达减少,对c-erbB-2的基因表达具有显著的反义抑制作用。

OBJECTIVE： This study was going to explore effects of c-erbB-2 antisense oligodeoxynucleotides （ASODN） conjugated epidermal growth factor （EGF） inhibited breast cancer cell SK-Br-3 in vitro. METHODS： c-erbB-2 ASODN, sense oligodeoxynucleotides （SODN） and nonsense oligodeoxynucleotides（NODN） were labeled by ^99mTc which then connected with EGF forming ^99mTc-ODN-EGF. Human breast cancer cell SK-Br-3 were incubated with ^99mTc-ODN-EGF, and then detected cells uptake percentage and refluent percentage. 5 × 104 cpm/d ^99mTc-ODN-EGF were added into cells incubating for 3 days, then detected cell vital force and c-erbB-2 protein expression. RESULTS： The cell uptake percentages of ^99mTc-ASODN-EGF, ^99mTc-SODN-EGF, ^99mTc-NODN-EGF and ^99mTc-ASODN 23. 70%,25. 24 %, 24. 91% and 5. 30% at 240 min. The cell uptake percentages of three EGF-drugs were evidently higher than ^99mTc-ASODN, P = 0.062. The cells refluent percentages of ^99mTc-ASODN-EGF,^99mTc-SODN-EGF and ^99mTc-NODN-EGF were 51.28% ,75. 70% and 81.20% at 12 h. The antisense oligodeoxynucleotide group was significantly fallen, P=0. 002. The cells vital force which were effected by four groups drugs were 0. 51 ± 0. 13,2. 33± 0. 48,2. 57± 57 and 2. 12±0. 45, respectively. The cell energy of antisense ODN was distinctly lower than other three groups, P = 0. 009,0. 014,0. 036. The c-erbB-2 protein expression of four groups drugs were 118. 77±5. 63,134. 16±3. 88,134. 76±1.76 and 131.24± 6. 66 ,respectively, and ^99mTc-ASODN-EGF group cells were evidently decreased, P = 0. 013. CONCLUSIONS： ^99mTc- ASODN-EGF has higher cell uptake percentage and lower cells refluent percentages. It significantly decreased breast cancer cell livability and c-erbB-2 protein expression. Therefore the ^99mTc-ASODN-EGF has remarkably antisense inhibition effect for c-erbB-2 gene expression.