[Inhibitory effect of paeoniflorin on CAL27 cells of tongue squamous cell carcinoma and its mechanism].

PURPOSE: To investigate the inhibitory effect of paeoniflorin(PF) with different concentrations on CAL27 cells of tongue squamous cell carcinoma in vitro and its possible mechanism. METHODS: CCK-8 technique and clone formation trial were used to detect the effect of PF on the proliferation and clone formation of CAL27 cells. Scratch test and Transwell method was used to detect the effects of PF on migration and invasion of CAL27 cells. Staining of Hoechst33342 was employed to evaluate the influence of PF on apoptosis of CAL27 cells, while Western blot was utilized to investigate the effect of PF on the expression of NF-κB pathway related proteins and EMT related proteins. The effect of PF on NO production in CAL27 cells was detected by nitric oxide detection kit. Statistical analysis was performed with SPSS 27.0 software package. RESULTS: PF inhibited proliferation, migration, and invasion of CAL27 cells in vitro in a concentration-dependent way. Moreover, PF caused apoptosis of CAL27 cells. PF impeded NF-κB pathway activity, decreased the expression of P-P65, further reduced the expression of iNOS and MMP-9, suppressed the production of NO, and concurrently inhibited Vimentin,promoted E-cadherin. CONCLUSIONS: Paeoniflorin inhibits the proliferation, migration and invasion of CAL27 cells, which may play an anti-cancer role by inhibiting the activation of NF-κB pathway and EMT.

microRNA-628 inhibits the proliferation of acute myeloid leukemia cells by directly targeting IGF-1R.

BACKGROUND: A variety of microRNAs (miRNAs) are aberrantly expressed in acute myeloid leukemia (AML), and these dysregulated miRNAs perform crucial roles in tumorigenesis and progression of AML. miR-628-3p (miR-628), one of the miRNAs dysregulated in multiple types of human cancers, exerts antitumor roles in different cancer types. However, no specific study has explored the expression pattern and role of miR-628 in AML. MATERIALS AND METHODS: In this study, RT-qPCR was performed to detect miR-628 expression in AML tissues and cell lines. CCK-8 assay, flow cytometry analysis and xenograft tumor experiment was carried out to determine the functions of miR-628 in AML cells. The possible mechanism underlying the activity of miR-628 in AML cells was also explored using a series of experiments. RESULTS: Our results revealed the downregulated expression of miR-628 in patients with AML and AML cell lines. Ectopic expression of miR-628 resulted in the inhibition of AML cell proliferation and induction of cell cycle arrest and apoptosis in vitro and attenuation of tumor growth in vivo. Insulin-like growth factor 1 receptor (IGF-1R) was identified as a direct target gene of miR-628 in AML cells. IGF-1R expression was upregulated in patients with AML and upregulation of IGF-1R expression inversely correlated with miR-628 level. Furthermore, IGF-1R knockdown imitated the tumor suppressive effect of miR-628 in AML cells. Restoration of IGF-1R expression abrogated the effects of miR-628 on the proliferation, cycle status, and apoptosis rate of AML cells. miR-628 inhibited the activation of phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K)/protein kinase B (Akt) pathway in AML cells both in vitro and in vivo through the inhibition of IGF-1R expression. CONCLUSION: Our results demonstrate that miR-628 exhibits antitumor effects in AML through the direct targeting of IGF-1R and regulation of PI3K/Akt pathway, suggestive of its potential role as a therapeutic target in patients with this aggressive hematological malignant tumor.

H2AFY is a novel fusion partner of MECOM in acute myeloid leukemia.

The MECOM gene encoding a zinc finger protein that functions as a transcription factor, was located on chromosome 3q26, and rearrangements of MECOM often cause its overexpression in acute myeloid leukemia (AML). We identified H2AFY as a novel fusion gene partner of MECOM in an elderly male AML patient with cryptic 3q26 rearrangement using the whole transcriptome sequencing, who carried out abnormal karyotype of 46,XY,t(3;5)(q27;q31),add(14)(p11). We validated the existence of the unreported H2AFY-MECOM fusion gene by RT-PCR and Sanger DNA sequencing, and detected mutations of NRAS and BCOR in this patient. In addition, we found abnormally elevated expression of MECOM in this patient by quantitative-polymerase chain reaction (RQ-PCR). Further research is needed to investigate functional characterizations of this novel fusion in the development of AML.

Primary distal femur T-cell lymphoma after allogeneic haematopoietic stem cell transplantation for chronic myeloid leukaemia: a rare case report and literature review.

Post-transplant lymphoproliferative disorders originating from T lymphocytes are a rare complication of allogeneic haematopoietic stem cell transplantation (allo-HSCT) that are not usually associated with Epstein-Barr virus infection. A male patient diagnosed at the age of 15 years with chronic myeloid leukaemia (in the chronic phase) was initially treated with oral hydroxyurea. The disease entered an accelerated phase when the patient was 22 years old. Complete remission was achieved after one course of homoharringtonine and cytarabine. The patient then underwent human leucocyte antigen-matched sibling donor allo-HSCT. Just over 6.5 years after the allo-HSCT, a second primary tumour was located in the distal femur and diagnosed as T-cell non-Hodgkin's lymphoma (stage IV, group B). This was treated with various chemotherapy and radiotherapy regimens, but the outcomes were poor and the disease progressed. The T-cell lymphoma invaded many sites, including the skeleton, spleen and skin, and the patient died within 8 months of the diagnosis. This current case report highlights the need for the early detection and prevention of subsequent primary malignancies after allo-HSCT.

Acute promyelocytic leukemia with a STAT5b-RARα fusion transcript defined by array-CGH, FISH, and RT-PCR.

Acute promyelocytic leukemia (APL) is characterized by the generation of the PML-RARα fusion transcript as a result of a reciprocal chromosomal rearrangement, t(15;17)(q22;q12), with breakpoints within the PML gene and the RARα gene. In a small proportion of APL cases, RARα is fused with a number of alternative partner genes. The signal transducer and activator of transcription 5 beta (STAT5b) is one of the variant partners. Here, we describe one rare case with all-trans retinoic acid (ATRA) -unresponsive APL characterized by the STAT5b-RARα fusion transcript. Morphology and immunophenotypic analyses indicated the typical features of APL; however, cytogenetic analysis exhibited a normal karyotype, and importantly, results of interphase fluorescence in situ hybridization (FISH) or reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated that PML-RARα expression was negative. FISH analysis with the RARα dual-color break-apart rearrangement probe indicated a submicroscopic deletion of the 3' end of one RARA gene. Indeed, the STAT5b-RARα fusion transcript was found in this case by array-based comparative genomic hybridization and nested RT-PCR. To the best of our knowledge, we report here only the sixth APL patient in the world with the STAT5b-RARα fusion transcript. Additional clinical studies concerning the prognosis, response to therapy, and pathogenesis of APL patients with STAT5b-RARα fusion are necessary.

[Bag3 gene expression in chronic lymphocytic leukemia and its association with patients' prognosis].

The aim of this study was to investigate bag3 gene expression in chronic lymphocytic leukemia (CLL)patients and its association with clinical prognosis. A total of 46 blood samples from untreated CLL patients were collected, SYBR Green-based real-time PCR was used to detect the bag3 mRNA expression, and its association with prognostic index was analyzed by statistical software. The results showed that the median values of bag3 level detected by real-time PCR in 46 CLL patients and normal controls were 0.021 (0.0007 - 1.124) and 0.0025 (0.0005 - 0.014) respectively, the former was significantly higher than the latter. The bag3 level in drug-resistant group was obviously higher as compared with the drug-responsive group. No association was found between bag3 expression and patient clinical baseline information (gender and age) as well as established prognostic factors (lymphocyte count, disease stage, IgVH mutation status, cytogenetics analysis and CD38, ZAP 70 expression). It is concluded that the bag3 expression in CLL patients is markedly higher than that in normal controls, while the high bag3 level in CML patients is probably related with drug resistance, but is not related with clinically established prognostic factors.

[Immunophenotypic analysis of Philadelphia chromosome positive acute lymphoblastic leukaemia in adults].

The aim of this study was to explore the immunophenotypic characteristics of Philadelphia chromosome positive (Ph(+)) acute lymphoblastic leukaemia (ALL) in adults and to evaluate their significance in predicting prognosis and guiding clinical treatment of diseases. The cell immunophenotypes of leukemic marrow or blood samples from 35 cases of Ph(+) ALL and 59 cases of Philadelphia chromosome negative (Ph(-)) ALL were detected by multiparameter flow cytometry, and their abnormal expressions were analysed. The results showed that the expression of all the Ph(+)ALL cases was found in B-cell lineage. As compared with Ph(-)B-ALL cases, the Ph(+)B-ALL cases displayed the higher expression of CD34 and CD13 (p < 0.05), but lower expression of CD38 (p < 0.05). The coexpressed rates of CD13, CD33 and CD15 in cases of Ph(+)B-ALL and Ph(-)B-ALL were 85.7% and 61.0% respectively. The former was higher than the later (p < 0.05). It is concluded that the Ph(+)ALL cases have the unique immunophenotype. The immunophenotypic analysis of CD34, CD13 and CD38 in adult B-ALL cases contributes to judging the existence of Ph chromosome. Thereby for adult ALL patients having above-mentioned unique immunophenotypes, the detection of bcr/abl fusion gene must be performed. Such phenotypic profile is helpful for predicting the poor outcome of the disease, and for defining patients who require different treatment strategies.