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Aim To investigate the effect of miR-320a up-regulation on the apoptosis and migration of Bel7402 cells induced by ribonucleic acid Ⅱ.Methods The different expression levels of miR-320a in normal liver cells and hepatocellular carcinoma (HCC) cells were detected by qRT-PCR.Bel-7402 cell was transfected with miR-320a mimic,and the miR-320a expression levels were measured by qRT-PCR.The effect of ribonucleic acid Ⅱ on proliferation of Bel-7402 and Bel-7402-miR-320a cells was measured by CCK-8 assay,and cell cycle and apoptosis were detected by flow cytometry.The migration and invasion ability of ribonucleic acid Ⅱ on Bel-7402 cells were tested by Transwell method.The expression of p53,Cyclin D1,Bax,Bcl-2,MMP-3 proteins were examined by Western blot.Results miR-320a expression levels in HCC cell line Bel-7402 were significantly lower than those in normal cell line HL-7702.Bel-7402 cells were successfully transfected with miR-320a mimic,named Bel-7402-miR-320a.CCK-8 showed that ribonucleic acid Ⅱ could effectively inhibit the proliferation of Bel7402 and Bel-7402-miR-320a cells in vitro in a dosedependent manner at the range of 100,200,300,400,500 mg · L-1.The IC50 of ribonucleic acid Ⅱexposure on Bel-7402 and Bel-7402-miR-320a cells for 12 h and 24 h was 250,200 mg · L-t and 150,120 mg · L-1,respectively;flow cytometric analysis indicated that over-expression of miR-320a could arrest Bel-7402 and Bel-7402-miR-320a cells induced by ribonucleic acid Ⅱ in G0/G1 phase,and promote the apoptosis of HCC cells.Transwell method showed that Bel-7402-miR-320a + Ribonucleic acid Ⅱ group could significantly inhibit the migration of HCC cells compared with control group and Bel-7402 + Ribonucleic acid Ⅱ group.Western blot results showed that the expression of p53,Bax proteins increased,while the Cyclin D1,Bcl-2,MMP-3 proteins were down-regulated in Bel-7402 and Bel-7402-miR-320a cells induced by ribonucleic acid Ⅱ.Conclusions The expression of miR-320a is lower in HCC cells than that in normal cell line.While ribonucleic acid Ⅱ could promote the apoptosis of liver cancer cells by arresting the cell cycle protein expression of Cyclin D1,activating p53 signaling pathway,down-regulating Bcl-2,up-regulating Bax and destroying Bcl-2/Bax proportions,and inhibiting the migration and invasion of HCC cells by downregulating MMP-3.Overexpression of miR-320a could increase the sensitivity and boost the pharmacological effects of ribonucleic acid Ⅱ on HCC cells.
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OBJECTIVE:To explore the effect of comprehensive intervention mode on the rational use of Ribonucleic acid Ⅱ for injection,and to provide reference for the management of adjuvant drugs for cancer therapy.METHODS:The rational use of ribonucleic acid Ⅱ was interfered by establishing evaluation criteria,reviewing medical record,establishing tumor therapy adjuvant management work group,classifying drug prescription right,examining and approving off-label drug use,strengthening the assessment and training,clinical pharmacists intervention.The utilization of ribonucleic acid Ⅱ was analyzed statistically in our hospital during Apr.-Jun.2015 (before intervention),Jul.-Sept.2015 (after the first intervention),Oct.-Dec.2015 (after the second intervention) and Jan.-Mar.2016 (after the third intervention).RESULTS:The reasonable rate of Ribonucleic acid Ⅱ for injection was 77.34％ before intervention,and 83.25％,83.64％,95.12％ after the first,second and third intervention respectively;the difference was statistically significant compared to before intervention (P＜0.05).The irrational types included inappropriate indications,unsuitable treatment course,inappropriate usage and dosage,and unsuitable drug combination,etc.The percentage of these irrational types decreased from 2.96％,4.93％,13.79％ and 0.99％ before intervention to 1.63％,0,3.25％ and 0 after the third intervention,respectively.The utilization rate of Ribonucleic acid Ⅱ for injection was reduced from 8.81％ before intervention to 3.93％ after the third intervention,the differences were statically significant (P＜0.05).CONCLUSIONS:The comprehensive intervention model combined with multiple intervention methods can promote the rational use of Ribonucleic acid Ⅱ for injection.It is suggested to further study and evaluate the intervention effect of this model on other adjuvant drugs for cancer therapy.
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Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .
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Objective:To establish a method for determining the immunological activity of ribonucleic acid. Methods: Leucocyte adherence inhibition test ( LAI) was applied, and the important parameters of LAI including the mouse strain, drug concentration, treatment time, content of buffer solution and cell density were researched. The immunological activity of RNAⅠ, Ⅱand Ⅲ was re-spectively determined by the method. Results:Stable and reliable parameters were obtained: the sample concentration was 10 mg· ml-1 , the treatment time was 2 hours, Ca2+ and Mg2+ were necessary for the buffer solution, and the cell density was about 4 × 107 cell·ml-1 . The strain of mouse showed no effect on the results. As a result, the determination method for immunological activity was established. Using the method, the immunological activity of RNA Ⅰ,Ⅱand Ⅲ was determined 3 times, and the results met the re-quirements with RSD below 20%. Conclusion:The method is suitable for determining the immunological activity of RNA.