胃肠病学和肝病学杂志
胃腸病學和肝病學雜誌
위장병학화간병학잡지
CHINESE JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY
2015年
4期
398-401
,共4页
李博%马俊%郭婉薇%陈晓武%王伟福%柳勤译%蔡少薇%叶丽芳%杨夏彤%王丽萍%许鸣
李博%馬俊%郭婉薇%陳曉武%王偉福%柳勤譯%蔡少薇%葉麗芳%楊夏彤%王麗萍%許鳴
리박%마준%곽완미%진효무%왕위복%류근역%채소미%협려방%양하동%왕려평%허명
微小RNA%胃癌%细胞周期%miR-345
微小RNA%胃癌%細胞週期%miR-345
미소RNA%위암%세포주기%miR-345
MicroRNA%Gastric cancer%Cell cycle%miR-345
目的：探讨miR-345影响胃癌细胞生长周期的作用机制。方法采用脂质体介导转染miR-345反义寡核苷酸至胃癌细胞MGC803、MKN45以沉默miR-345表达，采用实时英光定量PCR（Real-time PCR）、蛋白免疫印迹（Western blotting）检测胃癌细胞中p21 Cip1水平，随后检测CDK4、CDK6、Cyclin D1、p-Rb、Rb蛋白水平，进一步克隆形成实验比较其增殖能力，流式细胞术检测miR-345对胃癌细胞周期的影响。结果脂质体介导转染miR-345反义寡核苷酸至胃癌细胞MGC803、MKN45可以抑制miR-345的表达；p21Cip1的mRNA和蛋白水平在低miR-345表达的胃癌细胞系中升高；下调miR-345的表达后MGC803、MKN45细胞中CDK4、CDK6、Cyclin D1和总Rb蛋白的表达水平无明显变化，但Rb蛋白的磷酸化水平显著降低。结论沉默miR-345的表达能够明显阻滞胃癌细胞从G1期向S期的进程，减慢细胞生长速度；其机制与上调p21 Cip1进而抑制Rb蛋白的磷酸化有关。
目的：探討miR-345影響胃癌細胞生長週期的作用機製。方法採用脂質體介導轉染miR-345反義寡覈苷痠至胃癌細胞MGC803、MKN45以沉默miR-345錶達，採用實時英光定量PCR（Real-time PCR）、蛋白免疫印跡（Western blotting）檢測胃癌細胞中p21 Cip1水平，隨後檢測CDK4、CDK6、Cyclin D1、p-Rb、Rb蛋白水平，進一步剋隆形成實驗比較其增殖能力，流式細胞術檢測miR-345對胃癌細胞週期的影響。結果脂質體介導轉染miR-345反義寡覈苷痠至胃癌細胞MGC803、MKN45可以抑製miR-345的錶達；p21Cip1的mRNA和蛋白水平在低miR-345錶達的胃癌細胞繫中升高；下調miR-345的錶達後MGC803、MKN45細胞中CDK4、CDK6、Cyclin D1和總Rb蛋白的錶達水平無明顯變化，但Rb蛋白的燐痠化水平顯著降低。結論沉默miR-345的錶達能夠明顯阻滯胃癌細胞從G1期嚮S期的進程，減慢細胞生長速度；其機製與上調p21 Cip1進而抑製Rb蛋白的燐痠化有關。
목적：탐토miR-345영향위암세포생장주기적작용궤제。방법채용지질체개도전염miR-345반의과핵감산지위암세포MGC803、MKN45이침묵miR-345표체，채용실시영광정량PCR（Real-time PCR）、단백면역인적（Western blotting）검측위암세포중p21 Cip1수평，수후검측CDK4、CDK6、Cyclin D1、p-Rb、Rb단백수평，진일보극륭형성실험비교기증식능력，류식세포술검측miR-345대위암세포주기적영향。결과지질체개도전염miR-345반의과핵감산지위암세포MGC803、MKN45가이억제miR-345적표체；p21Cip1적mRNA화단백수평재저miR-345표체적위암세포계중승고；하조miR-345적표체후MGC803、MKN45세포중CDK4、CDK6、Cyclin D1화총Rb단백적표체수평무명현변화，단Rb단백적린산화수평현저강저。결론침묵miR-345적표체능구명현조체위암세포종G1기향S기적진정，감만세포생장속도；기궤제여상조p21 Cip1진이억제Rb단백적린산화유관。
Objective To investigate the the effect of miR-345 in the growth cycle of gastric cancer cell .Methods miR-345 antisense oligonucleotides were transfected into MGC 803, MKN45 cells.Real-time PCR was conducted to de-tect the expression of miR-345 in transfected cells.The expression of p21Cip1, CDK4, CDK6, Cyclin D1, p-Rb, Rb were examined by Western blotting and Real-time PCR.Then the cell cycle progression was examined by flow cytome-try.Results The expression of miR-345 in MGC803, MKN45 cells transfected with antisense miR-345 was significantly reduced.miR-345 knockdown could up-regulate the expression of p21Cip1 and the level of p-Rb was significantly re-duced, but no obvious change in the levels of the CDK 4, CDK6, Cyclin D1, Rb.The cell proliferation activity of MGC803, MKN45 cells was inhibited.The cell cycle was arrested in G1 phase.Conclusion miR-345 knockdown can obviously block the process of gastric cancer cells from G 1 phase to S phase .Its mechanism is associated with up-regula-ting p21 Cip1 and suppressing Rb protein phosphorylation .
