中华胰腺病杂志
中華胰腺病雜誌
중화이선병잡지
CHINESE JOURNAL OF PANCREATOLOGY
2010年
3期
199-201
,共3页
范钰%吴莺%徐军%钟锡明
範鈺%吳鶯%徐軍%鐘錫明
범옥%오앵%서군%종석명
胰腺肿瘤%畸胎瘤衍生生长因子-1%肿瘤侵袭%小分子干扰RNA
胰腺腫瘤%畸胎瘤衍生生長因子-1%腫瘤侵襲%小分子榦擾RNA
이선종류%기태류연생생장인자-1%종류침습%소분자간우RNA
Pancreatic neoplasms%Teratocarcinoma-derived growth factor-1%Neoplasm invasiveness%Small interfering RNA
目的 探讨TDGF-1基因沉默对胰腺癌细胞株PANC1侵袭力的影响.方法 设计并合成3个(S1、S2、S3)靶向TDGF-1基因的小干扰RNA(small interfering RNA,siRNA),筛选出沉默效果最好的siRNA.以该siRNA的不同浓度(3.125、6.25、12.5 nmol/L)转染PANC1细胞,并设对照组和脂质体组.采用实时定量PCR和Western blotting检测TDGF-1 mRNA和蛋白表达,以软琼脂集落培养试验和Boyden小室检测癌细胞的锚着不依赖性增殖和侵袭能力,并将转染48 h的细胞接种裸鼠,观察癌细胞的体内侵袭.结果 siRNA转染组细胞的TDGF-1 mRNA和蛋白表达水平呈浓度和时间依赖性下降,且较脂质体组明显降低(P值均＜0.01).对照组细胞集落形成数和穿膜细胞数分别为19.8±2.2和49.8±2.6;siRNA组呈浓度依赖性明显减少,12.5 nmol/L转染组分别为5.6±1.2和8.1±1.1.对照组、脂质体组和12.5 nmol/L转染组接种4周后裸鼠种植瘤体积分别为(2.228±0.026)cm3、(2.186±0.028)cm3和(0.728±0.023)cm3.对照组和脂质体组种植瘤侵犯周围肌层组织,转染细胞组未见侵犯.结论 采用RNA干扰技术沉默PANC 1细胞的TDGF-1基因表达可抑制癌细胞的侵袭力.
目的 探討TDGF-1基因沉默對胰腺癌細胞株PANC1侵襲力的影響.方法 設計併閤成3箇(S1、S2、S3)靶嚮TDGF-1基因的小榦擾RNA(small interfering RNA,siRNA),篩選齣沉默效果最好的siRNA.以該siRNA的不同濃度(3.125、6.25、12.5 nmol/L)轉染PANC1細胞,併設對照組和脂質體組.採用實時定量PCR和Western blotting檢測TDGF-1 mRNA和蛋白錶達,以軟瓊脂集落培養試驗和Boyden小室檢測癌細胞的錨著不依賴性增殖和侵襲能力,併將轉染48 h的細胞接種裸鼠,觀察癌細胞的體內侵襲.結果 siRNA轉染組細胞的TDGF-1 mRNA和蛋白錶達水平呈濃度和時間依賴性下降,且較脂質體組明顯降低(P值均＜0.01).對照組細胞集落形成數和穿膜細胞數分彆為19.8±2.2和49.8±2.6;siRNA組呈濃度依賴性明顯減少,12.5 nmol/L轉染組分彆為5.6±1.2和8.1±1.1.對照組、脂質體組和12.5 nmol/L轉染組接種4週後裸鼠種植瘤體積分彆為(2.228±0.026)cm3、(2.186±0.028)cm3和(0.728±0.023)cm3.對照組和脂質體組種植瘤侵犯週圍肌層組織,轉染細胞組未見侵犯.結論 採用RNA榦擾技術沉默PANC 1細胞的TDGF-1基因錶達可抑製癌細胞的侵襲力.
목적 탐토TDGF-1기인침묵대이선암세포주PANC1침습력적영향.방법 설계병합성3개(S1、S2、S3)파향TDGF-1기인적소간우RNA(small interfering RNA,siRNA),사선출침묵효과최호적siRNA.이해siRNA적불동농도(3.125、6.25、12.5 nmol/L)전염PANC1세포,병설대조조화지질체조.채용실시정량PCR화Western blotting검측TDGF-1 mRNA화단백표체,이연경지집락배양시험화Boyden소실검측암세포적묘착불의뢰성증식화침습능력,병장전염48 h적세포접충라서,관찰암세포적체내침습.결과 siRNA전염조세포적TDGF-1 mRNA화단백표체수평정농도화시간의뢰성하강,차교지질체조명현강저(P치균＜0.01).대조조세포집락형성수화천막세포수분별위19.8±2.2화49.8±2.6;siRNA조정농도의뢰성명현감소,12.5 nmol/L전염조분별위5.6±1.2화8.1±1.1.대조조、지질체조화12.5 nmol/L전염조접충4주후라서충식류체적분별위(2.228±0.026)cm3、(2.186±0.028)cm3화(0.728±0.023)cm3.대조조화지질체조충식류침범주위기층조직,전염세포조미견침범.결론 채용RNA간우기술침묵PANC 1세포적TDGF-1기인표체가억제암세포적침습력.
Objective To investigate the effects of the silence of teratocarcinoma-derived growth factor-1 ( TDGF-1 ) gene on invasion of human pancreatic cancer cell. Methods Three small interfering RNA (siRNA) targeting for TDGF-1 genes (S1, S2, S3 ) were designed and established, then the gene with the best silencing effects was screened. Human pancreatic cancer cell line PANC1 were transfected by siRNA with different concentrations (3. 125, 6.25, 12.5 nmoL/L), the cells without transfection, and simply treated with liposomes were controls. The expressions of mRNA and protein of TDGF-1 were determined by real time PCR and Western blot assay, respectively. The anchorage-independent growth was examined by clon formation in soft agar, and invasion ability was evaluated by boyden chamber model. PANC1 cells with transfection for 48h were injected into the nude mice to evaluate the invasion ability in vivo. Results The expressions of TDGF-1 mRNA and protein of cells transfected by siRNA were decreased in a dose-and time-dependent manner, which were significantly lower than those in liposomes group. Number of colony formation and transmembrane cell were 19.8 ± 2.2 and 49.8 + 2.6 in the control group, and 5.6 + 1.2 and 8. 1 + 1.1 in the 12.5 nmol/L transfection group. The volumes of tumor 4 weeks after transplation in the control group, liposomes group and the 12.5 nmol/L transfection group were (2.228 ± 0.016 ) cm3, ( 2.186 ± 0.028 )cm3 and ( 0.728 ± 0.023 )cm3. Conclusions TDGF-1 gene silence could inhibit invasion ability of human pancreatic cancer cell PANC1.
