中华消化外科杂志
中華消化外科雜誌
중화소화외과잡지
CHINESE JOURNAL OF DIGESTIVE SURGERY
2008年
3期
196-199
,共4页
黄小兵%李靖%梁平%郑璐%刘世呈%韩克强
黃小兵%李靖%樑平%鄭璐%劉世呈%韓剋彊
황소병%리정%량평%정로%류세정%한극강
BC047440基因%RNA干扰%肝肿瘤
BC047440基因%RNA榦擾%肝腫瘤
BC047440기인%RNA간우%간종류
BC047440 gene%RNA interference%Liver cancer
目的 构建shRNA并干扰肝癌HepG2细胞中BC047440基因,观察其对肝癌HepG2细胞增殖的影响.方法 根据BC047440基因序列,设计和合成两条特异表达该基因shRNA序列并克隆载体pGenesil-1,转染肝癌HepG2细胞并用定量PCR观察抑制BC047440基因表达的效果,并通过MTT法和流式细胞仪检测细胞增殖及细胞周期的变化.结果 酶切和序列测定提示成功构建了特异表达BC047440基因的shRNA1和shRNA2两个质粒.shRNA1和shRNA2对BC047440 mRNA的抑制率分别为80.22%和58.63%.干扰BC047440基因后,肝癌HepG2细胞增殖受到明显抑制,主要停滞在S期.结论 构建的shRNA能有效干扰肝癌HepG2细胞BC047440基因的表达,并抑制肝癌HepG2细胞的增殖,提示BC047440基因和肝癌HepG2细胞增殖有关.
目的 構建shRNA併榦擾肝癌HepG2細胞中BC047440基因,觀察其對肝癌HepG2細胞增殖的影響.方法 根據BC047440基因序列,設計和閤成兩條特異錶達該基因shRNA序列併剋隆載體pGenesil-1,轉染肝癌HepG2細胞併用定量PCR觀察抑製BC047440基因錶達的效果,併通過MTT法和流式細胞儀檢測細胞增殖及細胞週期的變化.結果 酶切和序列測定提示成功構建瞭特異錶達BC047440基因的shRNA1和shRNA2兩箇質粒.shRNA1和shRNA2對BC047440 mRNA的抑製率分彆為80.22%和58.63%.榦擾BC047440基因後,肝癌HepG2細胞增殖受到明顯抑製,主要停滯在S期.結論 構建的shRNA能有效榦擾肝癌HepG2細胞BC047440基因的錶達,併抑製肝癌HepG2細胞的增殖,提示BC047440基因和肝癌HepG2細胞增殖有關.
목적 구건shRNA병간우간암HepG2세포중BC047440기인,관찰기대간암HepG2세포증식적영향.방법 근거BC047440기인서렬,설계화합성량조특이표체해기인shRNA서렬병극륭재체pGenesil-1,전염간암HepG2세포병용정량PCR관찰억제BC047440기인표체적효과,병통과MTT법화류식세포의검측세포증식급세포주기적변화.결과 매절화서렬측정제시성공구건료특이표체BC047440기인적shRNA1화shRNA2량개질립.shRNA1화shRNA2대BC047440 mRNA적억제솔분별위80.22%화58.63%.간우BC047440기인후,간암HepG2세포증식수도명현억제,주요정체재S기.결론 구건적shRNA능유효간우간암HepG2세포BC047440기인적표체,병억제간암HepG2세포적증식,제시BC047440기인화간암HepG2세포증식유관.
Objective To investigate whether the proliferation of HepG2 ceils is influenced by interfering BC047440 gene with small hairpin RNA (shRNA) expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA (shRNA1 and shRNA2) were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells.
