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Objective:To investigate the effect and potential mechanism of miR-199a-5p on the radiosensitivity of cervical cancer CaSki cells.Methods:Cervical cancer CaSki cells were cultured in vitro. MiR-199a-5p mimics (miR-199a-5p mimics) were transfected into cervical cancer CaSki cells (miR-199a-5p group) with liposome by Lipofectamine 2000. CaSki cells transfected with mimics control were used as negative control (NC group) and non transfected CaSki cells were used as blank control (control group). After X-ray irradiation, real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-199a-5p in cells of each group. The effects of miR-199a-5p on radiosensitivity and apoptosis of CaSki cells were detected by clone formation assay and flow cytometry apoptosis assay. Bioinformatics software was used to predict the target gene of miR-199a-5p. Double luciferase reporter gene assay and Western blot were used to verify the targeting relationship between miR-199a-5p and thyroid hormone receptor interaction factor 4 (TRIP4). Results:The expression of miR-199a-5p in miR-199a-5p group was significantly higher than that in control group ( P<0.05). After X-ray irradiation, the expression of miR-199a-5p was more obvious ( P<0.05); Overexpression of miR-199a-5p could reduce the clonogenic ability and promote the apoptosis of CaSki cells ( P<0.05); Overexpression of miR-199a-5p could further reduce the clonal formation and promote the apoptosis of irradiated cells ( P<0.05); Double luciferase reporter gene experiment and Western blot confirmed that miR-199a-5p could target and negatively regulate TRIP4. Conclusions:miR-199a-5p can increase the radiosensitivity of cervical cancer CaSki cells by negatively regulating the expression of TRIP4.
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Objective:To investigate the effect and mechanism of miRNA-5193 (miR-5193) on the sensitivity of cervical cancer Caski cells to cisplatin.Methods:The expression of miR-5193 in cervical cancer cell lines C33A, SiHa, Caski and normal cervical cell line Ect1/E6E7 were determined by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). Caski cells were divided into control group (no transfection, normally cultured), miR-5193-negative control (miR-NC) group (transfected with miR-NC mimic), miR-5193 group (transfected with miR-5193 mimic), miR-NC+cisplatin group (transfected with miR-NC mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin group (transfected with miR-5193 mimic and treated with 10 μg/ml cisplatin), miR-5193+cisplatin+NC group (cotransfected with Foxp3-negative control vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin), and miR-5193+cisplatin+Foxp3 group (cotransfected with Foxp3 overexpression vector and miR-5193 mimic, and treated with 10 μg/ml cisplatin). Proliferation was detected by methyl thiazolyl tetrazolium (MTT), cell cycle was detected by PI single staining method, cell apoptosis was detected by Annexin V-FITC/PI double staining method, and expressions of CDK2, p27 and C-caspase-3 proteins in cells were detected by Western blot. Bioinformatics software was used to predict miR-5193 target genes, and the luciferase reporting system was used to identify the targeting relationship.Results:The relative expression of miR-5193 in cervical cancer C33A, SiHa and Caski cells was lower than that in normal cervical Ect1/E6E7 cells (0.56±0.06, 0.41±0.03, 0.23±0.02 vs. 1.00±0.10, all P < 0.05). Compared with the control group and miR-NC group, the cell proliferation activity (absorbance value) in miR-5193, miR-NC+cisplatin and miR-5193+cisplatin groups decreased (0.58±0.06, 0.59±0.07 vs. 0.38±0.04, 0.40±0.05, 0.23±0.02, all P < 0.05), the cell apoptosis rate increased [(2.5±0.2)%, (2.7±0.3)% vs. (12.6±1.2)%, (11.9±1.5)% , (18.9±1.7)%, all P < 0.05], and the proportion of cells in G 0/G 1 phase increased [(50.4±4.2)%, (51.3±6.3)% vs. (62.3±3.2)%, (61.9±5.8)%, (71.4±5.4)%, all P < 0.05]. The expression levels of p27 and C-caspase-3 proteins increased, and the expression level of CDK2 protein decreased. The software predicted that the target gene of miR-5193 was Foxp3, which was confirmed by the luciferase reporting system. Compared with the miR-5193+cisplatin+NC group, the cell proliferation activity (absorbance value) in miR-5193+ cisplatin+Foxp3 group increased (0.24±0.03 vs. 0.65±0.05, t = 21.094, P < 0.01), the proportion of cells in G 0/G 1 phase decreased [(71.0±6.4)% vs. (60.3±4.1)%, t = 4.196, P < 0.01], the apoptosis rate of cells decreased [(19.6±1.6)% vs. (11.5±1.2)%, t = 11.880, P < 0.01], the expression levels of p27 and C-caspase-3 proteins in cells decreased, and the expression levels of CDK2 and Foxp3 proteins increased. Conclusion:The miR-5193 may increase the sensitivity of cervical cancer Caski cells to cisplatin in vitro by targeted inhibition of the Foxp3 gene.
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@#Nanocrystals are nanoscale (1-1000 nm) dispersion systems in which small numbers of surfactants or polymers are used as stabilizers to disperse insoluble drug particles in water or oil. Nanocrystals enjoy not only high drug content, but also a simple and mature preparation process. At present, 24 nanocrystals products that have been marketed mainly focus on enhancing the solubility and bioavailability of poorly soluble drugs. And recent years have witnessed an increasing number of research reports on target drug delivery of nanocrystals through particle size control and surface modification. This paper mainly introduces three targeting strategies for prolonging the in vivo circulation time of nanocrystals, increasing the affinity for tumor cells and achieving the response to internal and external stimuli, and discusses the current challenges in the application of nanocrystal technology to targeted anti-tumor drugs.
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OBJECTIVE@#To screen the microRNAs (miRNAs) targeting Rictor and investigate their effects in regulating the biological behaviors of colorectal cancer (CRC).@*METHODS@#Human colorectal cancer cell line KM12SM was transfected with the miRNAs targeting Rictor identified by prediction software to test inhibitory effects of these miRNAs on Rictor expression using qRT-PCR and Western blotting. Dual luciferase reporter assay was used to further confirm the binding of these miRNAs to the 3'UTR of Rictor mRNA. Cell survival and colony formation assays were used to investigate the effects of these miRNAs on survival and colony formation in KM12SM cells.@*RESULTS@#miR-152 and miR-448 were identified as the Rictor-targeting miRNAs, which significantly inhibited the expression of Rictor in KM12SM cells ( < 0.05). The two miRNAs were confirmed to bind to the 3'UTR of Rictor mRNA and significantly inhibited luciferase activity in KM12SM cells ( < 0.01, < 0.05); they also showed activities of posttranscriptional modulation of Rictor. Overexpression of miR-152 and miR-448 both significantly inhibited the growth and colony formation of KM12SM cells.@*CONCLUSIONS@#miR-152 and miR-448 can down-regulate the protein expression of Rictor by targeting Rictor mRNA to negatively regulate the growth and colony formation of colorectal cancer cells.
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Humans , 3' Untranslated Regions , Cell Line, Tumor , Cell Proliferation , Colorectal Neoplasms , Drug Therapy , Gene Expression Regulation, Neoplastic , MicroRNAs , Pharmacology , Rapamycin-Insensitive Companion of mTOR ProteinABSTRACT
Objective To explore the efficacy and safety of apatinib combined with S-1 in patients with advanced NSCLC without sensitive gene mutation or unknown mutation status.Methods One hundred and four patients with advanced NSCLC without sensitive gene mutation or unknown mutation status were selected from the oncology department of the First Hospital of Qinhuangdao City,Hebei Province from April 2015 to April 2017.All patients refused intravenous chemotherapy.One hundred and four patients were randomly divided into treatment group (apatinib combined with S-1 group) and control group (S-1 alone group) by 1:1 digital method.However,two patients in the treatment group transferred to the control group for personal reasons.There is 50 cases in apatinib combined with S-1 group and 54 cases in S-1 group.The efficacy and adverse reactions of the two groups were evaluated.Results The objective remission rate was 48.0％ (24/50) and 27.8％ (15/54) (x2=4.530,P =0.033),the disease control rate was 82.0％ (41/50) and 74.1％ (40/54) (x2=0.947,P=0.331),the median PFS was 6.6 months and 3.4 months (t=25.555,P =0.000),the median OS was 16.0 months and 10.5 months (t =59.439,P =0.000),respectively.The overall incidence of adverse reactions was 82.0％ (41/50) and 70.4％ (38/54) respectively (x2 =1.923,P=0.166),of which 18.0％ (9/50) and 13.0％ (7/54) were more than grade 3 respectively (x2 =0.506,P =0.477).There was no death caused by treatment-related adverse reactions in both groups.Conclusion Appatinib combined with S-1 capsule has good short-term and long-term efficacy in the treatment of advanced non-small cell lung cancer without gene mutation or unknown mutation.The adverse reactions are tolerable and can be used as first-line treatment for patients unwilling to receive intravenous chemotherapy.
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Objective@#To investigate the effect of 2, 2', 4, 4'-tetrabromodiphenyl ether (PBDE-47) on the mitochondrial mass in rat adrenal pheochromocytoma (PC12) cells and the potential mechanisms.@*Methods@#Highly differentiated PC12 cells were divided into control, 1, 10 or 20 μmol/L PBDE-47-treated groups and cultured for 24 h. Transmission electron microscopy was employed to observe the changes in mitochondrial morphology and quantity in PC12 cells. Flow cytometry was used to measure the fluorescence intensity of Nonyl Acridine Orange (NAO) , a fluorescent indicator of mitochondrial membrane cardiolipin, to reflect mitochondria mass. Western blotting was used to determine the expression levels of Mitofusion 1 (Mfn1) and Fission 1 (Fis1) proteins. To further explore the role of abnormal mitochondrial fusion and fission in PBDE-47-induced mitochondrial mass changes, PC12 cells were divided into control group, 5 μmol/L M1 treatment group, 20 μmol/L PBDE-47 treatment group and 5 μmol/L M1+20 μmol/L PBDE-47 combined treatment group and cultured for 24 h, then the fluorescence intensity of NAO and expression levels of Mfn1 and Fis1 proteins were detected.@*Results@#The control group showed numerous mitochondria with normal morphology, while the number of mitochondria decreased after PBDE-47 treatment. Especially, the disappeared cristae, swelling and vacuoles of mitochondria and decreased fluorescence intensity of NAO (P<0.05) were observed in 10 and 20 μmol/L PBDE-47-treated groups. Meanwhile, the expression levels of Mfn1 and Fis1 proteins in the 10 and 20 μmol/L PBDE-47-treated groups were significantly decreased compared with control group (P<0.05) . However, 5 μmol/L M1 co-treatment with 20 μmol/L PBDE-47 significantly increased the levels of Mfn1 and Fis1 proteins and fluorescence intensity of NAO compared with the 20 μmol/L PBDE-47 group (P<0.05) .@*Conclusion@#PBDE-47 can inhibit the mitochondrial fusion and fission process, thus leading to damage of mitochondria mass in PC12 cells.
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Objective To evaluate the diagnostic value of the level of plasma D-dimer,high-density lipoprotein (HDL),carcino embryonic antigen (CEA) and carbohydrate antigen 724 (CA724) in gastric cancer.Methods The plasma and clinicopathological data of 103 gastric cancer patients and 111 normal controls were collected.The levels of D-dimer,HDL,CEA and CA724 were detected.SPSS 13.0 statistical software was applied to analysis the sensitivity and specificity of each examination method and to find out the appropriate combination.Results The levels of D-dimer,CEA and CA724 in patients with gastric carcinoma were 0.87 (2.69) μg/ml,2.66 (4.38) ng/ml,5.10 (7.79) U/ml,respectively,they were distinctly higher than those in normal controls [0.22 (0.21) μg/ml,1.28 (1.60) ng/ml,1.81 (1.60) U/ml,all P =0.000].HDL level was significantly lower in patients than that in normal controls [0.86 (0.35) mmol/L vs 1.29 (0.44) mmol/L,P=0.000].The area ofROC curve of D-dimer,HDL,CEA,CA724 were 0.799,0.859,0.739,0.743,respectively.The cut-off of D-dimer was 0.46 μ.g/ml,the sensitivity was 68.0 ％,the specificity was 86.5 ％.The cut-off of HDL was 0.995 mmol/L,the sensitivity was 73.8 ％,the specificity was 84.7 ％.The cut-off of CEA was 3.585 ng/ml,the sensitivity was 44.7 ％,the specificity was 92.0 ％.The cut-off of CA724 was 3.765 U/ml,the sensitivity was 57.3 ％,the specificity was 89.2 ％.The sensitivity of D-dimer+HDL+CA724 was 83.5 ％,the specificity was 89.2 ％.The sensitivity and specificity of D-dimer+HDL+CEA+CA724 were 84.5 ％ and 89.2 ％,respectively.Conclusions The D-dimer+HDL+CEA+CA724 may provide the evidence for diagnosis of gastric cancer.Combined detection has higher sensitivity and specificity.
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Objective To investigate the significance of plasma D-dimers,high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol (LDL-C) and apolipoprotein a (LP (a)) of patients with gastrointestinal cancer.Methods One hundred and thirty-nine cases with gastrointestinal cancer and 155 healthy persons were selected as subjects.Latex enhanced immunoassays method was used to measure the D dimer concentration.Direct method was applied to detect the levels of serum HDL-C,LDL-C,LP (a)Chemiluminescence method was applied to detect serum carcinoembryonic antigen (CEA)concentration.Determine the sensitivity and specificity of each index in gastrointestinal cancer,and explore the relationship between the concentration and the lymph node metastasis and distant metastasis.Results The levels of D-dimer,LDL-C,LP (a),CEA in patients with gastrointestinal carcinoma were 0.80 (2.37) mg/L,3.46 (1.46) mmol/L,317 (262) mg/L,2.61 (3.62) μ,g/L respectively,significant higher than those of health control(0.21 (0.22) mg/L,2.86 (0.98) mtmol/L,139 (187) g/L,1.29 (1.42) μg/L respectively; Z =-8.388,-6.150,-8.589,-7.142,P ＜0.001).The level of HDL-C in gastrointestinal cancer patients was 0.86(0.38) mmoL/L,lower than that of health control(1.29(1.42) mmol/L; Z =-10.643,P ＜0.001)The area of ROC curve of D-dimmer,HDL-C,LDL-C,LP (a),CEA were 0.783,0.859,0.708,0.790,0.741 respectively.Compared with area of ROC curve of CEA,that of D-dimer,LDL-C,LP(a) were significant different (Z =1.110,0.809,1.257 ; P =0.27,0.42,0.21).Compared with CEA AUC,HDL-C AUC was significant different (Z =3.225,P =0.0013).Compared with patients with no lymph node metastasis,the levels of D-dimers,LDL-C,LP (a) in patients with lymph node metastasis were higher (P =0.003,0.002,0.005 respectively).And the concentration of HDL-C decreased significantly (P =0.001).Compared with patients without metastases,serum D-dimer,LDL-CLP (a) concentration in patients with distant metastasis were significant higher(P ＜ 0.001) and the concentration of HDL-C decreased (P ＜ 0.001).Conclusion The levels of D-dimer and lipoprotein might be proved the base for diagnosis or assessment of gastrointestinal malignant tumor.
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Objective To evaluate the efficacy and safety of radiation combined with targeted therapy of EGFR-TKI in the patients with stage IV non-small cell lung cancer ( NSCLC). Methods There were 17 female and 9 male patients with NSCLC enrolled into this study, which included 19 adenocarcinoma, 4 alveolar carcinoma and 3 uncertain carcinoma according to the iconography findings. Sixteen patients suffered from single or multiple bone metastasis,and 10 cases with brain metastasis. Gefitinib 250 mg or Erlotinib ISO mg per day were administrated during and after the process of radiation until the disease progressed. Results All patients had complete combined therapy, 12 of them suffered from diarrhea, 8 from emesia and 12 from erythra. The average score of ECOG improved from 3 to 2 after combined therapy. The bone metastasis control rate was 93.8% ,brain metastasis control rate was 70.0% , and the 6-month local lung lumps control rate was 84. 6%. Conclusion Palliative radiation combined with targeted therapy of EGFR-TKI is an effect and safe therapy for the patients with the stage IV of NSCLC, but the influence on survival shall be observed in further study.
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A total of 101 patients undergoing operations for malignant gastrointestinal tumors (stage Ⅱ to Ⅲ) were enrolled and randomly assigned to receive intraperitoneal hyperthermal chemotherapy plus intravenous chemical injection (treatment group, n=51) or routine intravenous chemical injection (control group, n=50). Our results indicated that the recurrence rate and the metastatic rate in the treatment group were significantly lower than those in the control group (25.5% vs. 50.0%, 13.7% vs. 30.0%, both P< 0. 05), although the 3-and 5 year-survival rates were significantly higher (both P < 0. 05). Our data suggest that intraperitaneal hyperthermal chemotherapy plus general chemotherapy after surgery for malignant gastrointestinal tumors could effectively reduce tumor recurrence and metastases and improve long-term survival.
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Sixty-one patients with moderate to severe malignant ascites were enrolled and randomly assigned to receive intraperitoneal hyperthermal chemotherapy+intravenous chemical injection (treatment group; n=31) or routine intravenous chemical injection (control group; n=30). Short-term response and reverse effects were observed. Our results indicated that the complete remission rate, part remission rate,and clinical benefit rate in the treatment group was significantly higher than those in the control group (38.71% vs 13.33% ,41.94% vs 16. 67%, and 90.32% vs 66.67%, respectively). No difference in reverse effects was found between the two groups. Our data suggest that intraperitoneal hyperthermal chemotherapy plus general chemotherapy may effectively control the malignant ascites, and the reverse effects might be well tolerated.