中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2012年
3期
175-179
,共5页
陶春华%陈腾飞%Praveen K.Yadav%邬瑞金%邱骅婧%吴维%刘占举
陶春華%陳騰飛%Praveen K.Yadav%鄔瑞金%邱驊婧%吳維%劉佔舉
도춘화%진등비%Praveen K.Yadav%오서금%구화청%오유%류점거
胃肿瘤%肽核酸类%寡核苷酸类,反义%粘蛋白类%肿瘤细胞,培养%肿瘤浸润
胃腫瘤%肽覈痠類%寡覈苷痠類,反義%粘蛋白類%腫瘤細胞,培養%腫瘤浸潤
위종류%태핵산류%과핵감산류,반의%점단백류%종류세포,배양%종류침윤
Stomach neoplasms%Peptide nucleic acids%Oligonucleotides,antisense%Mucins%Tumorcells cultured%Neoplasm invasiveness
目的 构建粘蛋白1 (MUC1)基因的反义肽核酸(PNA),观察对胃癌细胞MKN-45侵袭能力的影响,并探讨其机制.方法 根据MUC1基因的序列设计反义PNA序列,经脂质体介导转染胃癌细胞MKN45,并设空载体组(随机对照)和空白对照组(阴性对照)运用荧光定量PCR检测MUC1的表达,并观察E-粘附素的表达变化；Transwell小室实验观察对胃癌细胞侵袭力的影响.结果 构建的3个MUC1基因的反义PNA均能有效抑制该基因表达,其表达水平分别为0.62±0.18,0.49±0.12和0.60±0.21,显著低于阴性对照组(1.18±0.03,P＜0.01).阴性对照组与随机对照组之间差异无统计学意义(1.00±0.04,P=0.657).取抑制效率最高的PNA进行后续研究转染MUC1 PNA后的胃癌细胞侵袭能力显著下降(t=2.09,P=0.005),并伴有E-粘附素基因及蛋白的表达上调.结论 胃癌细胞MKN-45中MUC1基因与E-粘附素的表达存在负相关,抑制MUC1基因的表达能显著抑制肿瘤细胞的侵袭能力.
目的 構建粘蛋白1 (MUC1)基因的反義肽覈痠(PNA),觀察對胃癌細胞MKN-45侵襲能力的影響,併探討其機製.方法 根據MUC1基因的序列設計反義PNA序列,經脂質體介導轉染胃癌細胞MKN45,併設空載體組(隨機對照)和空白對照組(陰性對照)運用熒光定量PCR檢測MUC1的錶達,併觀察E-粘附素的錶達變化；Transwell小室實驗觀察對胃癌細胞侵襲力的影響.結果 構建的3箇MUC1基因的反義PNA均能有效抑製該基因錶達,其錶達水平分彆為0.62±0.18,0.49±0.12和0.60±0.21,顯著低于陰性對照組(1.18±0.03,P＜0.01).陰性對照組與隨機對照組之間差異無統計學意義(1.00±0.04,P=0.657).取抑製效率最高的PNA進行後續研究轉染MUC1 PNA後的胃癌細胞侵襲能力顯著下降(t=2.09,P=0.005),併伴有E-粘附素基因及蛋白的錶達上調.結論 胃癌細胞MKN-45中MUC1基因與E-粘附素的錶達存在負相關,抑製MUC1基因的錶達能顯著抑製腫瘤細胞的侵襲能力.
목적 구건점단백1 (MUC1)기인적반의태핵산(PNA),관찰대위암세포MKN-45침습능력적영향,병탐토기궤제.방법 근거MUC1기인적서렬설계반의PNA서렬,경지질체개도전염위암세포MKN45,병설공재체조(수궤대조)화공백대조조(음성대조)운용형광정량PCR검측MUC1적표체,병관찰E-점부소적표체변화；Transwell소실실험관찰대위암세포침습력적영향.결과 구건적3개MUC1기인적반의PNA균능유효억제해기인표체,기표체수평분별위0.62±0.18,0.49±0.12화0.60±0.21,현저저우음성대조조(1.18±0.03,P＜0.01).음성대조조여수궤대조조지간차이무통계학의의(1.00±0.04,P=0.657).취억제효솔최고적PNA진행후속연구전염MUC1 PNA후적위암세포침습능력현저하강(t=2.09,P=0.005),병반유E-점부소기인급단백적표체상조.결론 위암세포MKN-45중MUC1기인여E-점부소적표체존재부상관,억제MUC1기인적표체능현저억제종류세포적침습능력.
Objective To create Mucin gene 1 (MUC1) antisense peptide nucleic acid (PNA),and to observe its effects on MKN-45 cell invasion and explore the mechanism. Methods The sequence of antisense PNA was designed according to MUC1 gene sequence and transfected into human gastric cancer cells (MKN-45) by liposome,and the empty vector group (randomized control group)and blank control group (negative control group) were involved. The expression of MUC1 was detected by real time quantitative PCR and the changes of E-cadherin expression were also observed.The effects on gastric cancer cell invasion were tested with transwell chamber assays.Results The expression of MUC1 gene was effectively suppressed by the 3 created antisense PNA,and their expression level (0.62±0.18,0.49±0.12 and 0.60±0.21) was significantly lower than that of negative control group (1.18 ± 0.03,P ＜ 0.01). There was no significant difference between radomized control group and negative control group (1.00±0.04,P=0.657).After MUC1 PNA transfected,the capability of gastric cancer cell invasion decreased significantly (P=0.005).And the expression of E-cadherin at mRNA and protein level was up-regulated.Conclusions There is negative correlation between MUC1 and E-cadherin expression in gastric cancer cell MKN-45.The capability of tumor cell invasion is significantly inhibited by suppressing MUC1 gene expression.
