河北医药
河北醫藥
하북의약
HEBEI MEDICAL JOURNAL
2015年
17期
2579-2582
,共4页
胰腺癌%原癌基因表皮生长因子%环氧合酶2%基因沉默%增殖%凋亡%侵袭
胰腺癌%原癌基因錶皮生長因子%環氧閤酶2%基因沉默%增殖%凋亡%侵襲
이선암%원암기인표피생장인자%배양합매2%기인침묵%증식%조망%침습
pancreatic cancer%proto-oncogene epidermal growth factor%cyclooxygease 2%gene silencing%proliferation%apoptosis%invasion
目的：观察siRNA沉默胰腺癌BXPC-3细胞中Her-2／neu基因对COX-2表达及细胞增殖、凋亡、侵袭力的影响，探讨干扰Her-2／neu抑制BXPC-3生物学行为的可能机制，为胰腺癌的临床治疗提供新的思路。方法采用实时荧光定量PCR法（ RT-QPCR）和Western blot检测Her-2／neu、COX-2 mRNA和蛋白在胰腺癌BXPC-3细胞及正常胰腺HPDE6C7细胞的表达。设计并构建Her-2／neu siRNA慢病毒表达载体转染BXPC-3细胞，检测Her-2／neu基因沉默对COX-2 mRNA和蛋白表达的影响。 CCK-8法检测Her-2／neu siRNA对细胞增殖的影响。流式细胞仪检测Her-2／neu siRNA对细胞凋亡的影响。 Transwell小室实验检测Her-2／neu siRNA对细胞侵袭力的影响。结果 Her-2／neu、COX-2 mRNA和蛋白在胰腺癌细胞中均高表达，而在正常胰腺细胞株中极少表达，差异有统计学意义（ P ＜0换．05）。Her-2／neu基因沉默能显著抑制COX-2 mRNA和蛋白表达，细胞增殖、侵袭能力显著减弱，细胞凋亡率显著增加（ P ＜0．05）。结论胰腺癌细胞高表达Her-2／neu、COX-2，Her-2／neu基因沉默能够显著抑制细胞增殖和侵袭能力，并促进细胞凋亡。
目的：觀察siRNA沉默胰腺癌BXPC-3細胞中Her-2／neu基因對COX-2錶達及細胞增殖、凋亡、侵襲力的影響，探討榦擾Her-2／neu抑製BXPC-3生物學行為的可能機製，為胰腺癌的臨床治療提供新的思路。方法採用實時熒光定量PCR法（ RT-QPCR）和Western blot檢測Her-2／neu、COX-2 mRNA和蛋白在胰腺癌BXPC-3細胞及正常胰腺HPDE6C7細胞的錶達。設計併構建Her-2／neu siRNA慢病毒錶達載體轉染BXPC-3細胞，檢測Her-2／neu基因沉默對COX-2 mRNA和蛋白錶達的影響。 CCK-8法檢測Her-2／neu siRNA對細胞增殖的影響。流式細胞儀檢測Her-2／neu siRNA對細胞凋亡的影響。 Transwell小室實驗檢測Her-2／neu siRNA對細胞侵襲力的影響。結果 Her-2／neu、COX-2 mRNA和蛋白在胰腺癌細胞中均高錶達，而在正常胰腺細胞株中極少錶達，差異有統計學意義（ P ＜0換．05）。Her-2／neu基因沉默能顯著抑製COX-2 mRNA和蛋白錶達，細胞增殖、侵襲能力顯著減弱，細胞凋亡率顯著增加（ P ＜0．05）。結論胰腺癌細胞高錶達Her-2／neu、COX-2，Her-2／neu基因沉默能夠顯著抑製細胞增殖和侵襲能力，併促進細胞凋亡。
목적：관찰siRNA침묵이선암BXPC-3세포중Her-2／neu기인대COX-2표체급세포증식、조망、침습력적영향，탐토간우Her-2／neu억제BXPC-3생물학행위적가능궤제，위이선암적림상치료제공신적사로。방법채용실시형광정량PCR법（ RT-QPCR）화Western blot검측Her-2／neu、COX-2 mRNA화단백재이선암BXPC-3세포급정상이선HPDE6C7세포적표체。설계병구건Her-2／neu siRNA만병독표체재체전염BXPC-3세포，검측Her-2／neu기인침묵대COX-2 mRNA화단백표체적영향。 CCK-8법검측Her-2／neu siRNA대세포증식적영향。류식세포의검측Her-2／neu siRNA대세포조망적영향。 Transwell소실실험검측Her-2／neu siRNA대세포침습력적영향。결과 Her-2／neu、COX-2 mRNA화단백재이선암세포중균고표체，이재정상이선세포주중겁소표체，차이유통계학의의（ P ＜0환．05）。Her-2／neu기인침묵능현저억제COX-2 mRNA화단백표체，세포증식、침습능력현저감약，세포조망솔현저증가（ P ＜0．05）。결론이선암세포고표체Her-2／neu、COX-2，Her-2／neu기인침묵능구현저억제세포증식화침습능력，병촉진세포조망。
Objective To investigate the specific effect of silencing of Her-2/neu gene by small interfering RNA ( siRNA) on COX-2 expression and the cell biological behaviors of human pancreatic cancer cell line BXPC-3 in order to find an new therapeutic method for pancreatic cancer.Methods The expression levels of Her-2/neu, COX-2 mRNA and proteins were detected by RT-QPCR and Western Blot in human pancreatic cancer cell lines BXPC3 and normal pancreas cell line HPDE6C7.The specific Her-2/neu siRNA was synthesized and transfected into BXPC3 cells by lentivirus, and the expression levels of Her-2/neu and COX-2 were detected.The CCK-8 assay was used to analyze the proliferation of BXPC3 cells.Flow cytometry was used to determine the apoptosis of BXPC3 cells.The cell invasion was evaluated by transwell assay.Results The overexpressions of Her-2/neu, COX-2 mRNA and proteins were observed in human pancreatic cancer cell lines BXPC3. However the expressions of Her-2/neu, COX-2 mRNA and proteins were almost not detected in normal pancreas cell line HPDE6C7, there were significant differences between the two kinds of cells ( P <0.05).The Her-2/neu siRNA could remarkedly inhibit the expressions of COX-2 mRNA and proteins.The proliferation and invasiveness of BXPC3 cells in Her-2/neu siRNA group were decreased obviously.However the apoptosis of BXPC3 cells in Her-2n/eu siRNA group was significantly increased (P <0.05).Conclusion The overexpressions of Her-2/neu and COX-2 are observed in the human pancreatic cancer cell lines BXPC3,moreover, Her-2/neu gene silence can obviously inhibit proliferative and invasive abilities of the cells, furthermore, which can induce apoptosis of BXPC3 cells.
