ABSTRACT
ObjectiveTo observe the apoptotic effect of cardamonin on K562 cells and its relationship with the expressions of PTEN, p-Akt, NF-κB and Bcl-2.Methods K562 cells were treated with cardamonin for 48 h, and the following tests were performed:(1) The cell morphology was observed by light microscopy.(2)IC50 of the K562 cells was dtermined by MTT test.(3) The apoptosis rate was detected by flow cytometry.(4) The expressions of Bcl-2 and Bax mRNA were detected a by RT-PCR.(5) The expressions of PTEN, p-Akt, NF-κB and Bcl-2 proteins were detected by Western blot.Results Obvious apoptosis was observed in the K562 cells after treated with cardamonin for 48 h.MTT assay indicated that the proliferation of K562 cells was obviously inhibited in a dose-and time-dependent manner. Comparing with the blank group, the early apoptosis rate and expression of Bax mRNA were significantly increased.At the same time, the expression of Bcl-2 mRNA was significantly decreased.All of them presented a dose-dependent manner. The expression of PTEN obviously increased with the increasing dose of cardamonin and the expressions of p-Akt, NF-κB and Bcl-2 were decreased.Conclusions Cardamonin promotes the apoptosis in K562 cells in a dose-dependent manner by increasing the expression of PTEN and decreasing the expressions of p-Akt, NF-κB, and Bcl-2.
ABSTRACT
Objective: To study rDNA ITS (internal transcribed spacers) sequences variation from di erent population of Aquilaria sinensis in main habitat of China. Methods: The rDNA ITS regions of various A. sinensis were ampli ed by PCR method and sequenced, and they were analyzed by means of the software of CLUSTAL and MEGA. Results: The sequences of rDNA ITS region of A. sinensis were reported for the rst time, and the sequences of ITS region were 680bp (ITS1 246bp, 5.8S 163bp, ITS2 271bp). There were 6 variable sites among populations and the genetic distance were 0.0% to 1.1%,which indicated the intraspecific genetic variation was low of A. sinensis. Conclusion: The variation of rDNA ITS sequences can be used to authenticate A.sinensis from di erent geographical regions and their adulterants.
ABSTRACT
Objective To construct a cDNA library of three-year old Polygonum multiflorum leaf tissues so as to further research the gene regulation of secondary metabolite biosynthesis of medicinal plants. Methods Total RNA from leaf tissues of P.multiflorum was extracted and mRNA was purified,which were synthesized to double strand cDNA through reverse transcription.After the cDNA termini was blunted,the 5' end of EcoR Ⅰ adapters phosphorylated was conjoined,and then digested by Xho Ⅰ,cDNA fragments were fractionated by Sepharose CL-2B spin column.The fragments longer than 400 bp were linked to Uni-ZAP XR vector.The primary cDNA library was established after the recombinants had been packaged.Uni-ZAP XR Vector might fleetly release pBluescript SK-phasmids at the presence of ExAssist helper phage of coinfection and inverted E.coli SOLR.Finally,PCR and double enzymes digestion were used to analyze the range of inserts,respectively. Results The titer of cDNA primary library was 1.07?106pfu/mL and the length of exogenous insert was at about 0.5-2.0 kb with 5.4?105 recombinants,the recombinants of amplified library were 4.25?1011 and the rate of recombination was 98.5%. Conclusion The results indicate that the cDNA library of P.multiflorum leaf tissues has enough volume for screening the desired genes and sets up a basis for studying on gene regulation of secondary metabolite biosynthesis of medicinal plants besides.