ABSTRACT
Objective To establish a stable lung cancer A549 cell line transfected by AMP-activated protein kinase (AMPK) expression vector,and to observe the effect of AMPK on proliferation as well as on the invasive ability of A549 cells.Methods Full-length of AMPK gene was amplified and its target gene was digested,then inserted into the GV358 plasmid.Co-tranfected 293T cells were subjected to the lentivirus equipment package.Subsequently,we collected the lentivirus supernatant to infect the A549 cells and establish a stably,overexpressed cell line A549.The mRNA and protein of AMPK were examined by real-time quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting.The proliferation and invasion abilities of A549 cells were detected by methyl thiazolyl thiazolium (MTT) and Transwell assay.Results GV358-AMPK lentivirus vectors was successfully constructed by restrictive enzyme digestion and plasmid sequencing.There were significantly increased expressions of AMPK protein (5.87 times,P =0.002) and mRNA (16.12 times,P ＜ 0.001) after transfected with GV358-AMPK compared with the Vector group.Meanwhile,AMPK overexpression showed significantly lower proliferation (the forth day:0.53 ± 0.03 vs.0.64 ±0.05,P=0.021;the fifth day:0.58 ± 0.04 vs.0.80 ± 0.07,P =0.002) and weaken invasive ability [(1.6±0.5) ×l05 vs.(3.4±0.3) ×105,P=0.004] ofA549 cells.Conclusion The lentiviralAMPK expression vector and its A549 cell line is successfully constructed.AMPK overexpression inhibits the proliferation and invasive ability of A549 cells.
ABSTRACT
OBJECTIVE@#To investigate the rare earth elements(REEs) contents and distribution characteristics in nasopharyngeal carcinoma( NPC) tissue in Gannan region.@*METHOD@#Thirty patients of NPC in Gannan region were included in this study. The REEs contents were measured by tandem mass spectrometer inductively coupled plasma(ICP-MS/MS) in 30 patients, and the REEs contents and distribution were analyzed.@*RESULT@#The average standard deviation value of REEs in lung cancer and normal lung tissues was the minimum mostly. Light REEs content was higher than the medium REEs, and medium REEs content was higher than the heavy REEs content. REEs contents changes in nasopharyngeal carcinoma were variable obviously, the absolute value of Nd, Ce, Pr, Gd and other light rare earth elements were variable widely. The degree of changes on Yb, Tb, Ho and other heavy rare earth elements were variable widely, and there was presence of Eu, Ce negative anomaly(δEu=0. 385 5, δCe= 0. 523 4).@*CONCLUSION@#The distribution characteristic of REEs contents in NPC patients is consistent with the parity distribution. With increasing atomic sequence, the content is decline wavy. Their distribution patterns were a lack of heavy REEs and enrichment of light REEs, and there was Eu , Ce negative anomaly.