CAJ | 학술논문

目的：探讨慢病毒系统介导的LKB1基因在子宫内膜癌HEC-1A细胞中的过表达，为进一步研究LKB1基因在子宫内膜癌的作用机制奠定基础。方法以PCR扩增LKB1克隆质粒获得全长cDNA，将LKB1 cDNA链接到慢病毒载体pWPI，构建慢病毒表达质粒LKB1/pWPI。通过与包装质粒pCMV-Dr8.74和pMD2.G共转染293T细胞进行病毒包装，用包装成功后的病毒液感染子宫内膜癌HEC-1A细胞，以荧光定量PCR、Western blot法检测HEC-1A细胞中LKB1的相对表达量。结果成功扩增LKB1全长cDNA和构建LKB1重组慢病毒表达载体LKB1/pWPI。转染包装293T细胞后能产生慢病毒颗粒并能有效感染靶细胞HEC-1A。转染后HEC-1A-LKB1-pWPI细胞中LKB1的表达率明显高于亲本细胞和空白对照细胞（P约0.01）。结论成功构建携带LKB1基因的慢病毒表达载体，包装病毒后能有效地感染子宫内膜癌HEC-1A细胞，为进一步探讨LKB1基因在子宫内膜癌中的生物学效应奠定了基础。
목적：탐토만병독계통개도적LKB1기인재자궁내막암HEC-1A세포중적과표체，위진일보연구LKB1기인재자궁내막암적작용궤제전정기출。방법이PCR확증LKB1극륭질립획득전장cDNA，장LKB1 cDNA련접도만병독재체pWPI，구건만병독표체질립LKB1/pWPI。통과여포장질립pCMV-Dr8.74화pMD2.G공전염293T세포진행병독포장，용포장성공후적병독액감염자궁내막암HEC-1A세포，이형광정량PCR、Western blot법검측HEC-1A세포중LKB1적상대표체량。결과성공확증LKB1전장cDNA화구건LKB1중조만병독표체재체LKB1/pWPI。전염포장293T세포후능산생만병독과립병능유효감염파세포HEC-1A。전염후HEC-1A-LKB1-pWPI세포중LKB1적표체솔명현고우친본세포화공백대조세포（P약0.01）。결론성공구건휴대LKB1기인적만병독표체재체，포장병독후능유효지감염자궁내막암HEC-1A세포，위진일보탐토LKB1기인재자궁내막암중적생물학효응전정료기출。
Objective To investigate lentivirus-mediated LKB1 gene overexpression in HEC-1A endometrial cancer cells as a basis for future research. Methods The LKB1 gene was subcloned by RT-PCR from a cDNA plasmid into a lentiviral pWPI expression vector,generating the plasmid LKB1/pWPI.This plasmid was co-transfected into 293T cells together with the packaging plasmid pCMV-Dr8.74 and pMD2.G.The recombinant virus was used to infect HEC-1A cells,and LKB1 expression was measured using real-time quantitative PCR (qRT-PCR) and Western blotting. Results We succeeded in constructing the recombinant plasmid LKB1/pWPI and packaging it into lentiviral particles in 293T cells.This virus infected HEC-1A cells efficiently,leading to higher LKB1 expression than in the parental cells or untransfected controls (P<0.01). Conclusion The LKB1 gene has been incorporated into a lentiviral expression vector,allowing studies of its effects on the development of endometrial cancer.