1.miR-10b overexpressionpromotes the proliferation of lung cancer cell line A549
Jing TANG ; Zhongquan SHI ; Ling HU ; Guoli LI ; Xiaoling FENG ; Feng LIU
Basic & Clinical Medicine 2017;37(8):1108-1112
Objective To explore the of miR-10b cordribution in patients with non-small cell lung cancer (NSCLC) and adjacent tissues,and to investigate the effect of miR-10b on the malignant change of lung cancer cell A549 by regulating the expression of Kruppel-like factor 4 (KLF4).Methods Fourty patients with NSCLC were selected with lung cancer and in miR-10b expression adjacent tissues of lung cancer cell A549 transfected miR-10b mimics,changes of CCK-8 assay were used to detect the proliferation of lung cancer cells;real time PCR and Western blot were used to examine the cell KLF4 mRNA and protein levels;soft agar colony formation assay was used to detect the expression of miR-10b on the proliferation of lung cancer cell A549 tumor malignant.Results miR-10b expression in lung cancer A549 cells and lung cancer tissue were higher than that of normal lung epithelial 16HBE cells and cancer adjacent tissues;overexpression of miR-10banalogue in A549 cells,KLF4 protein levels significantly decreased,KLF4 mRNA was not changed significantly;miR-10b expression significantly increased the growth of A549 cells.Conclusions The distribution of miR-10b in different cell types and tissues may be different,which may promote the proliferation and malignancy of lung cancer cells by inhibitingthe expression of KLF4.
2.Effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis by inhibiting AKT, ERK and upstream signal SRC activity.
Shi-mei QI ; Jun LV ; Yu MENG ; Zhi-lin QI ; Lie-feng LING
China Journal of Chinese Materia Medica 2015;40(16):3267-3272
To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic
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pharmacology
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Apoptosis
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drug effects
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Breast Neoplasms
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drug therapy
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enzymology
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genetics
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physiopathology
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Down-Regulation
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Extracellular Signal-Regulated MAP Kinases
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genetics
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metabolism
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Female
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Humans
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MCF-7 Cells
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Proto-Oncogene Proteins c-akt
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genetics
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metabolism
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Saponins
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pharmacology
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Signal Transduction
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drug effects
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Triterpenes
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pharmacology
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src-Family Kinases
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genetics
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metabolism
3.Purification of the Anti-PMOP Effective Fraction from Camellia semiserrata Seed with DiaionHP-20 Macroporous Adsorption Resin
Hongjing LIN ; Xiaojuan WU ; Qian TANG ; Haifeng SHI ; Ling TANG ; Baomin FENG ; Yongqi WANG
China Pharmacy 1991;0(03):-
OBJECTIVE:To study the technical conditions and parameters of DiaionHP-20 macroporous adsorption resin for the purification of the anti-PMOP(primary Type I osteoporosis) active fraction from Camellia semiserrata seed.METHODS: With the elution ratios and purity of total flavonoids and polyphenol from Camellia semiserrata as indexes,the technical parameters of DiaionHP-20 macroporous adsorption resin including the adsorption quantity,the loading sample,the elution concentration and volume of alcohol were investigated.RESULTS: The adsorption quantity of DiaionHP-20 macroporous resin on total flavonoids and polyphenol were 59.5 mg?g-1 and 23.9 mg?g-1 respectively.The optimal technical conditions were as follows:the quantity of drug was 10.5 times that of resin;water-soluble foreign substance was washed off with distilled water weighing 3 times the weight of resin,and total flavonoids and polyphenol were eluted using 25% ethanol weighing 13.5 times the weight of resin.CONCLUSION:DiaionHP-20 macroporous resin had good purification effects on total flavonoids and polyphenol from Camellia semiserrata seeds.The process is simple,low cost,and suitable for lavge-scale production.
4.The regulatory activity of the lately cloned promoter PLUNC-p in transgenic Xenopus Laevis
Ling ZHANG ; Xiangling FENG ; Wen ZHOU ; Hong LI ; Jianling SHI ; Kaitai YAO
Chinese Journal of Pathophysiology 2000;0(08):-
AIM: Our study focused on investigation of tissue specific regulatory activity of the newly cloned promoter: PLUNC-p with driving enhanced green fluorescent protein ( EGFP ). METHODS: Transgenic Xenopus Laevis system was applied.RESULTS: The green fluorescence protein directed under PLUNC-p was expressed strictly in branchial arches and epidermis of Xenopus Laevis embryos while CMV promoter showed ubiquitous regulation characteristic.CONCLUSION: PLUNC-p is able to direct epithelia specific expression of EGFP . This property of PLUNC-p might raise the possibility that lead target genes to express tissue-specifically.
5.The influence of the life quality by the cognitive,behavioral,psychological intervention to the spouses of breast cancer patients receiving chemotherapy
Xiuying TANG ; Feng LUO ; Guo SHI ; Pengjuan JIN ; Ling ZHANG ; Hongyuan LI
Chongqing Medicine 2014;(22):2848-2849,2852
Objective To explore the influence of life quality for the breast cancer patients receiving chemotherapy after the cog-nitive ,behavioral and psychological intervention to their spouse .Methods 120 breast cancer patients received standardized chemo-therapy and their spouses ,and divided into control and intervention groups .The intervention group receive the routine care and health guidance .Before and after chemotherapy ,the life quality of patients was investigated .The data was analyzed statistically .Re-sults The result by the breast cancer patients Quality of Life Questionnaire in Chinese (FACT-B) show that ,the scores of the con-trol and intervention groups in the physiological status ,social/family status ,emotional status ,functional status ,additional attention andtotalscorewere(18.77±4.18,16.48±4.60,17.35±4.41,16.04±4.80,20.81±6.02,89.45±6.34 ;22.46±3.57,19.03± 4 .83 ,18 .58 ± 3 .96 ,18 .59 ± 4 .48 ,24 .73 ± 5 .63 ,103 .39 ± 8 .91) .The scores of intervention groups was increased significantly than the control group .The data was analyzed statistically .Conclusion The quality of life of patients was improved by the cognitive ,be-havioral and psychological guidance and intervention to the spouse of breast cancer chemotherapy patients.
6.Molecular Cloning and Sequencing of HCV E2 Gene
Dong-wei, ZHANG ; Bu-feng, LIANG ; Zi-bai, QI ; Shi-Gan, LING
Virologica Sinica 2001;16(1):40-44
HCV RNA positive serum was first selected by RT-PCR test kit from several anti-HCV positive sera obtained from Xi'an.HCV RNA extrac ted from the elected sera was converted to cDNA by reverse transcription with ra ndom primer.Half-nested PCR was performed.The amplified product was 852 bp.The purified PCR product was digested by restriction endonucleases and then ligated to epressio vector pET-22b\++.Its nucleotide sequence was determined by dideoxy chain termination method.A comparison of the sequence with several isolates rep orted previously showed that the sequence belonged to HCV type Ⅱ.
7.Side population cells in human gallbladder cancer cell line GBC-SD regulated by TGF-β-induced epithelial-mesenchymal transition.
Zhifa, ZHANG ; Feng, ZHU ; Ling, XIAO ; Min, WANG ; Rui, TIAN ; Chengjian, SHI ; Renyi, QIN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(6):749-55
Mounting evidence has shown that side population (SP) cells are enriched for cancer stem cells (CSCs) responsible for cancer malignancy. In this study, SP technology was used to isolate a small subpopulation of SP cells in human gallbladder cancer cell line GBC-SD, and SP cells which had superior potential for proliferation in vitro and tumorigenesis in vivo were identified. Importantly, the abundance of GBC-SD SP cells was increased by a transforming growth factor-β (TGF-β)-induced epithelial-mesenchymal transition (EMT), and this effect was accompanied with a strong up-regulation of ABCG2 mRNA expression, and a decreased sensitivity to mitoxantrone. SP cells were restored upon the removal of TGF-β and the reversion of the cells to an epithelial phenotype, and smad3-specific siRNA reduced SP abundance in response to TGF-β. In conclusion, TGF-β-induced EMT by smad-dependent signaling pathway promotes cancer development and anti-cancer drug resistant phenotype by augmenting the abundance of GBC-SD SP cells, and a better understanding of mechanisms involved in TGF-β-induced EMT may provide a novel strategy for preventing cancer progression.
8.Preliminary analysis of miRNA expression profile of chemosensitivity for TPF regimen in hypopharyngeal squamous cell carcinoma
Haizhou WANG ; Meng LIAN ; Ru WANG ; Jie ZHAI ; Ling FENG ; Qian SHI ; Hongzhi MA ; Jugao FANG
Chinese Archives of Otolaryngology-Head and Neck Surgery 2016;23(4):205-210
[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.
9.Dynamic Changes of Type Th22 Cell Immunological Response During Atherosclerosis Process in Experimental Mice
Jun WAN ; Lei SHI ; Qingwei JI ; Ying SHI ; Ling LIU ; Zhengde LU ; Ying FENG ; Jing YE ; Yingzhong LIN
Chinese Circulation Journal 2016;31(5):454-458
Objective: To study the dynamic changes of type Th22 cell immunological response during atherosclerosis process in experimental mice in order to provide a new theoretical basis for atherosclerosis therapy. Methods: 8 weeks C57BL/6J mice were divided into 2 groups: Experiment group,n=24 ApoE-/- mice and Control group, n=24 normal mice. All animals received high fat diet and the following indexes were compared between 2 groups at 0, 4, 8, 12 weeks after treatment: aortic atherosclerotic lesions were deifned by Oil red O staining, dynamic changes of Th22 cells in spleen were measured by lfow cytometry, mRNA expressions of interleukin-22 (IL-22), IL-22R1, AhR and T-bet in aorta were examined by RT-PCR, blood levels of IL-22 was detected by ELISA. Results: Compared with Control group, Experiment group had the increased area of aortic atherosclerosis (the ratio of plaque area/lumen area) and Th22 cell (CD4+ IL-22+/CD4+T cell) amount, elevated mRNA expressions of IL-22, IL-22R1, AHR, T-bet in aorta and higher blood levels of IL-22 at all time points, the differences between each time point (except 0 week) had the statistic meaning,P<0.05. In Experiment group, the differences between 2 adjacent time points, for the area of aortic atherosclerosis and mRNA expressions of AHR, T-bet: 4 weeks vs 0 week, 8 weeks vs 4 weeks, 12 weeks vs 8 weeks all had statistic meaning; for Th22 cell amount: 4 weeks vs 0 week, 8 weeks vs 4 weeks had statistic meaning and 12 weeks vs 8 weeks had no real distinction; for mRNA expressions of IL-22, IL-22R1 and blood levels of IL-22: 4 weeks vs 0 week had statistic meaning and 8 weeks vs 4 weeks, 12 weeks vs 8 weeks had no real distinctions. Conclusion: Hyperactive immunological response of Th22 cells might be involved in atherosclerosis process, the relevant mechanism should be further studied.
10.Protein interaction network analysis of Panax notoginseng saponins.
Zhen-Zhen REN ; Yan-Ling ZHANG ; Xing WANG ; Shi-Feng WANG ; Yu-Su HE ; Chen-Xi ZHAI ; Yan-Jiang QIAO
China Journal of Chinese Materia Medica 2014;39(11):2097-2101
Panax notoginseng (PN) is one of the commonly used clinical medicines for cardiovascular diseases and possesses a variety of pharmacological effects. P. notoginseng saponins (PNS) are the most important bioactive components in PN. The purpose of this study was to explain the mechanism of PNS on molecular network level. 18 targets of the main medicinal ingredients of PNS were gained by virtual screening based on pharmacophores and data mining. A protein interaction network of PNS was constructed with 189 nodes and 721 interactions. By a graph theoretic clustering algorithm Molecular Complex Detection (MCODE), 14 modules were detected. Gene ontology (GO) enrichment analysis of the modules demonstrated that the roles of PNS played in cardiovascular disease related to multiple biological processes, which could represent the characteristics of traditional Chinese medicine (TCM) as a whole to regulate the disease. The results showed that the blood circulation and hemostasis efficacy of PN related with the biological processes such as positive regulation of cAMP metabolic and biosynthetic process, platelet activation and regulation of blood vessel size, regulation of T cell proliferation and differentiation and so on. Therefore, the module-based network analysis will be an effective method for better understanding TCM.
Drugs, Chinese Herbal
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chemistry
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Humans
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Panax notoginseng
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chemistry
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Protein Interaction Maps
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drug effects
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Proteins
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chemistry
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Saponins
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chemistry