1.Effects of small interfering RNA on Survivin expression, proliferation and apoptosis of hepatocellular carcinoma cell line MHCC-97H
Zhaoxia ZHANG ; Ziqin LIU ; Yanfei CHEN ; Litian XUAN ; Tianyou WANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(9):707-710
Objective To observe the effect of small interfering RNA(siRNA) targeting Survivin gene on survivin expression,proliferation and apoptosis of hepatocellular carcinoma cell line MHCC-97H.Methods Survivin sequence specific siRNA was designed and synthesized.siRNA/liposome complex was transfected into hepatocellular carcinoma cell line MHCC-97H.The MHCC-97H cells were divided into Survivin siRNA group(Si-survivin),negative control siRNA group(NC group)and blank group (normal control group).Survivin mRNA and protein expressions were detected by reverse transcription-PCR and Western blot,respectively.The proliferation of MHCC-97H was measured by methythiazolydiphenyl-tetrazolium bromide assay.The Annexin V/PI double labeled flow cytometry was employed to measure the apoptosis at 24 h after transfection in different concentrations of Survivin siRNA(12.5,25.0 and 50.0 nmoL/L,respectively).Results After 48 h of transfection,the Survivin mRNA levels were 0.55 ± 0.16 (Si-survivin group),0.85 ± 0.28 (NC group) and 0.93 ± 0.40 (normal control group),respectively,which were significantly different among 3 groups (F =414,P < 0.01).The level of Survivin mRNA was the lowest in Si-survivin group,which was statistically different with NC group and normal control group (t =-20.56,-28.37,all P < 0.001).The levels of Survivin protein expression in 3 groups were 0.602 ± 0.005 (Si-survivin group),0.835 ± 0.007 (NC group) and 0.993 ± 0.003 (normal control group) at 48 h after transfection,which were statistically different among 3 groups (F =238,P <0.01).The lowest level of protein expression was in Si-survivin group,which was statistically different with NC group and normal control group (t =-40.17,-66.03,all P < 0.001).After 72 h and 96 h of transfection,the inhibitory rate of cell growth was significantly higher in Si-survivin group [(19.5 ± 3.6)%,(12.0 ± 0.9)%] compared with that in NC group [(3.6 ± 0.9) %,(-1.3 ± 6.1) %] (t--36.18,42.53,all P < 0.05).The apoptosis rates in 12.5,25.0,50.0 nmol/L Survivin siRNA were (22.64 ± 2.54) %,(35.37 ± 3.28) % and (53.28 ± 4.35) %,respectively.However,in NC group and normal control group,the apoptosis rates were (8.77 ± 1.25) % and (9.72 ± 1.37) %.The rates were statistically different among those 5 groups(F =35.93,P <0.01).And in the apoptosis rates of siRNA groups in different concentratiom were statistically different when compared between each two groups (t =-29.73,-38.57,all P < 0.001).Conclusion Survivin specific siRNA can inhibit the proliferation and induce the apoptosis by blocking Survivin gene expression in hepatocellular carcinoma cell line MHCC-97H.
2.Clinical analysis of two pediatric precursor B-cell acute lymphoblastic leukemia with C-MYC rearrangement
Lei ZHANG ; Rong LIU ; Juanjuan LI ; Dixiao ZHONG ; Mei YUE ; Litian XUAN
Chinese Journal of Applied Clinical Pediatrics 2021;36(9):687-690
Objective:To explore the characteristics, diagnosis, and treatment of precursor B-cell acute lymphocytic leukemia with C- MYC rearrangement (preBLL) in children. Methods:The clinical data in 2 cases of childhood preBLL in Department of Hematology, Children′s Hospital Affiliated to Capital Institute of Pediatrics in June and August 2019 were summarized and analyzed.Results:Both cases were acute lymphoblastic leukemia with precursor B-cell immunophenotype.Hepatosplenomegaly and peripheral white blood cells were significantly increased, and the morphology of bone marrow was L3. C- MYC rearrangement was discovered by cytogenetic tests.Both children have received the treatment of the mature B-cell tumor protocol (FAB/LMB96), and early remission was developed in 1 case with TP53 gene mutation but relapsed thereafter and died finally.Another case had reached sustained complete remission after treatment. Conclusions:Children with preBLL is rare, and routine C- MYC rearrangement should be performed in children with Precursor B-cell lymphoblastic leukemia whose morphology of bone marrow was L3.Its treatment needs to be further studied, and multi-center clinical trials need to be actively conducted to analyze and summarize large numbers of cases to identify effective protocol and improve the prognosis.
3.Advances in the treatment of hypereosinophilic syndromes
International Journal of Pediatrics 2020;47(11):764-767
Hypereosinophilic syndromes(HES)are a heterogeneous group of disorders characterized by persistent overproduction of eosinophils in bone marrow, peripheral blood, and tissues, which can involve multiple tissues and organs at the same time.Eosinophils infiltration and medium release can lead to multiple organ damage.The choice of treatment options depend on clinical manifestations, laboratory test results, and mutation analysis.The purpose of this review is to summarize the current treatment of HES, including conventional therapies such as glucocorticoids, hydroxylurea, interferon-alpha, as well as small molecule targeted drugs led by tyrosine kinase inhibitors, interleukin-5 and its receptor targeted monoclonal antibodies.
4.Study on silence of survivin gene in cancer cells A549, Hela S3 and K562 by small interfering RNA and its sequence screening
Mengze HU ; Xiaodong SHI ; Tianyou WANG ; Rong LIU ; Litian XUAN ; Yanfei CHEN
Chinese Journal of Applied Clinical Pediatrics 2014;29(3):194-198
Objective To investigate the biological response of survivin siRNA in A549 human lung cancer cells and Hela S3 human cervical cancer cells as well as K562 human erythroleukemia cells,in order to screen the working sequences of survivin siRNA.Methods Three sequences of survivin-targeted siRNA were designed and synthetized,and human cancer lines of A549,Hela S3,K562 were transfected with Hiperfect liposome entrapped survivin siRNA,respectively.The expression of survivin mRNA was detected by means of real-time polymerase chain reaction (RT-PCR) with SYBR Green Ⅰ.Cell proliferation was detected by way of WST-8 cell count kit at 48 hours and 72 hours after transfection.Results The expression of survivin mRNA in all 3 cancer cells studied was significantly inhibited by all siRNA at 48 hours and 72 hours after transfection,gene expression inhibition ratio were 57.47%-88.53% after transfection 48 hours and were 69.94%-95.03% after transfection 72 hours.Cell proliferation was also significantly inhibited 48 hours and 72 hours after transfection,cell multiplication inhibition ratio were 27.88%-47.36% after transfeotion 48 hours and were 42.59%-57.29% after transfeciton 72 hours.Sequence 1 had the most inhibition efficacy on survivin gene expression and proli-feration in tumor cells.Inhibition rate of the three tumor cell gene expression of survivin at 48 hours are above 75%,72 h all over 90%,48 hours of cell proliferation inhibition rate are above 40%,72 hours of more than 50%.Conclusions The chemo-synthesized siRNAs can significantly down-regulate survivin mRNA expression in cancer cell studies.Survivin siRNA is capable of inhibiting the proliferation of tumor cell in vitro and it might be a new strategy for tumor-targeted therapy.Sequence 1 is the most efficacious working survivin siR-NA in the study.