ABSTRACT
Objective To explore the impact of heat shock factor 2 (HSF2) on the development of lung cancer by promoting interleukin (IL)-10 expression.Methods 50 lung cancer tissues and adjacent normal tissues selected from 50 patients,the expression level of mRNA and protein of HSF2 and IL-10 were respectively detected by RT-PCR,Western blot and Immunohistochemistry;To interfere with expression of HSF2 in A549 cells by siRNA,the expression level of IL-10 was detected by Western blot.Results Compared with the adjacent normal tissues,HSF2 of 76％ (38 of 50) cases were up-regulated (P＜0.01),IL-10 of 80％ (40 of 50) eases were up-regulated (P＜0.01),protein level consistent with mRNA level.The up-regulation expression of IL-10 in lung cancer tissues and HSF2 positively correlated (R2 =0.921 6).The expression of IL-10 in A549 cells was weakened through interference with HSF2 by siRNA.Conclusion HSF2 may contribute to the development of lung cancer by facilitating the expression of IL-10.
ABSTRACT
In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.