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Objective: To detect the expression levels of microRNA200 (miR200) family in the plasma of the breast cancer patients and the normal controls, and to evaluate their potential values in the screening, progression and prognosis evaluation of breast cancer. Methods: A total of 82 cases of plasma samples of the patients with breast cancer (breast cancer group) and 30 cases of healthy plasma samples (control group) were collected. The microRNAs (miRNAs) were extracted from the plasma samples, the expression levels of miR200 family (miR200a, miR200b, miR200c, miR141 and miR429) were quantified by using real-time PCR technique. The receiver-operating characteristic (ROC) curve was used to assess the diagnostic value of miRNAs, and the associations between the levels of miRNAs and the clinicopathological characteristics were analyzed. Cox proportional hazard mode was used for survival analysis. Results: Compared with control group, the expression level of miR141 in the plasma of the patients in breast cancer group was decreased (P 0 . 05). Concuson: MiR200b and miR141 are likely to become the molecular biological parameters for the diagnosis of breast cancer.
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Objective To study the cytotoxicity of bi-chimeric antigen receptors modified T lymphocytes (BiCAR-T) on the human acute myeloid leukemia (AML) cell line HL60 in vitro and the anti-tumor effects of BiCAR-T on the NOD SCID mouse model of AML in vivo.Methods The BiCAR-T were prepared and the expression of chimeric antigen receptor (CAR) of prepared BiCAR-T was analyzed by flow cytometry.In vitro study was divided into two groups:the experiment group (BiCAR-T) and the control group (T lymphocyte).The killing rate of BiCAR-T in vitro on HL60 cells was determined by CCK8 assay and the level of interferon-γ (IFN-γ) secreted from BiCAR-T co-culturing with HL60 cells for 48 hours was detected by enzyme linked immunosorbent assay (ELISA) at different effect/target ratios (5∶1,10 ∶ 1,20 ∶ 1).The NOD SCID mice AML model was established by the injection of HL60 cells through tail vein and used to assess the antitumor effects in vivo.The mice were randomly divided into three groups according to the random number table:the blank control group receiving 0.9％ NaCl 0.2 ml through tail vein,the model group and the treatment group receiving 1 × 107 HL60 cells in 0.2 ml phosphate buffer saline (PBS).After 20 days,the treatment group was injected with 2 × 107BiCAR-T in 0.2 ml PBS 3 times a week for 2 weeks,while the other two groups received 0.9％ NaCl 0.2 ml.The pathological changes in the mice livers and spleens were observed after 2 weeks of treatment.Results The CAR expression rates of BiCAR-T were more than 50.00％.In vitro experiments proved that the killing rates of BiCAR-T in the experimental group and T lymphocytes in the control group on HL60 cells were (25.43 ±1.32)％ vs.(16.18 ±0.75)％,(50.33±3.11)％ vs.(25.47±1.27)％,and (85.89 ± 3.96) ％ vs.(49.45 ± 2.77) ％ at different effect/target ratios (5 ∶ 1,10 ∶ 1,20 ∶ 1).The killing efficiency of BiCAR-T and T lymphocytes on HL60 cells was significantly different (F =404.17,P ＜ 0.001);the killing efficiency of BiCAR-T and T lymphocytes on HL60 cells was significantly different at different effect/ target ratios (F =548.09,P ＜ 0.001);and the killing efficiency on HL60 cells in the experimental group (BiCAR-T) was significantly higher than that in the control group (T lymphocytes) at different effect/target ratios (F =45.36,P ＜ 0.001).The IFN-γlevels secreted from BiCAR-T in the experiment group and T lymphocytes in the control group co-culturing with HL60 ceils after 48 h were (435.65 ± 20.44) pg/ml vs.(356.75 ± 19.87) pg/ml,(1 639.98 ± 95.75) pg/ml vs.(1 109.37 ± 80.98) pg/ml,and (3 467.43 ± 187.54)pg/ml vs.(2 245.52 ± 112.66)pg/ml.The IFN-γlevel in the experiment group (BiCAR-T) and the control group (T lymphocytes) was significantly different (F =156.24,P ＜ 0.001);the IFN-γ level was significantly different at different effect/target ratios (F =857.67,P ＜ 0.001);the IFN-γlevel in the experimental group (BiCAR-T) was significantly higher than that in the control group (T lymphocytes) at different effect/ target ratios of 5 ∶ 1,10 ∶ 1,20 ∶ 1,respectively (F =46.31,P ＜ 0.001).The result of hematoxylineosin staining (HE) staining showed that leukocyte infiltration in the treatment group was significantly decreased compared with the model group.Conclusion The experimental results showed that BiCAR-T is a kind of efficient targeted immunocyte modified by gene engineering,and it can significantly inhibit leukocyte infiltration of AML in vivo and in vitro.
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OBJECTIVE:To discuss the effect of trastuzumab combined with chemotherapy on efficacy and serum tumor mark-ers in patients with human epidermal growth factor receptor 2(HER2)positive advanced gastric cancer. METHODS:74 advanced gastric cancer patients in HER2 positive were randomly divided into observation group and control group,37 cases in each group. Control group received cisplatin + capecitabine chemotherapy;based on it,observation group additionally received trastuzumab by intravenous infusion in the first day,8 mg/kg. 21 d was a course,and it lasted for 6 months. The short-term efficacy,long-term effi-cacy,serum tumor marker contents and toxic and side effects were compared between 2 groups. RESULTS:The effective rate (56.76% vs. 32.44%)and control rate(89.19% vs. 65.86%)in observation group were significantly higher than control group,the differences were statistically significant(P0.05). CONCLUSIONS:Trastuzumab combined with chemotherapy helps to reduce serum tumor markers content,and improve short-term efficacy,long-term efficacy of advanced gastric cancer patients with positive HER2.
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BACKGROUND:Studies have shown that microRNAs (miRNAs) have moderating effect on the renewal and differentiation of cancer stem cels. However, there is no complete understanding on the effect of microRNA-17-92 gene on gastric cancer stem cel renewal and proliferation. OBJECTIVE:To explore the effect of miRNA-17-92 in promoting self-renewal and proliferation of gastric cancer stem cels. METHODS:(1) The gradualy reduced miRNAs during gastric cancer stem cel self-renewal were investigated using miRNA array based on RNAs from differentiated and adherent cels. (2) The miRNA-17-92 was constructed and transfected to gastric cancer stem cels. (3) The effects of miRNA-17-92 on the self-renewal of gastric cancer stem cels were studied by tumor sphere assayin vitro. (4) The effects of miRNA-17-92 on the proliferation of gastric cancer stem cels were investigated by MTT assay and colony formation assay. RESULTS AND CONCLUSION:(1) miR-19b/20a/92a expression gradualy reduced in the adhesion and differentiation of gastric cancer stem cels. (2) The expression of lentivirus carrying miRNA-17-19 gene in MKN28 cels and CD44-/EpCAM- cels were significantly increased; transient transfection of pre-miR-19b/20a/92a increased the expression of CD44-/EpCAM- and MKN28 miRNA, transient transfection of pre-miR-19b/20a/92a antagonists reduced the expression of SGC7901 and CD44+/EpCAM+ miRNA; overexpression of lenti-miR-19b/20a/92a significantly increased the ability of gastric cels to form tumor spheres; chemotherapy drugs prolonged the survival time of lenti-miR-19b/20a/92a-infected cels; transient transfection of pre-miR-19b/20a/92a significantly increased the number of CD44+/EpCAM+ cels, but transfection of pre-miR-19b/20a/92a antagonist reduced the number of CD44+/EpCAM+ cels. (3) MTT proliferation assay showed that gastric cancer cel proliferation rate in miR-19b/20a/92a stably expressing group was faster than that in the control group. Transient transfection of miR-19b/20a/92a precursor accelerated the growth rate of gastric cancer cels, and transient transfection of its antagonist slowed down the growth rate of gastric cancer cels. Colony formation assay showed that transient transfection of miR-19b/20a/92a precursor significantly increased the colony formation number as compared with the control group; transient transfection of miR-19b/20a/92a antagonist reduced the colony formation as compared with the control group. These findings indicate that miR-19b/20a/92a gene presents with continuous deletion in gastric cancer stem cel differentiation process, and miRNA-17-92 gene can promote the renewal and proliferation of gastric cancer stem cels.