1.Construction of Peroxiredoxin 2 gene lentiviral expression vector and effect on colorectal cancer SW480 cells
Jihong FENG ; Zhongxue FU ; Kunming WEN ; Shouru ZHANG ; Weidong LU ; Hao WANG ; Xingye WU
Chinese Journal of Immunology 2014;(8):1078-1082
To construct the lentiviral vector containing Peroxiredoxin 2(Prdx2) gene and the colorectal cancer cell line stably transduced with Prdx 2-containing vector , so as to provide a useful tool for studying the role of Prdx 2 in colorectal cancer.Methods: Prdx2 was amplified by PCR and inserted into lentiviral expression vector Ubi-MCS-EGFP-IRES-Puromycin (GV218) to generate Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector.The inserted Prdx2 gene was verified by double enzyme digestion and DNA sequencing.Subsequently ,lentiviruses were produced and transduced into SW 480 cells.EGFP expression was examined under fluorescence microscopy ,the expression of Prdx2 was detected with qRT-PCR and Western blot.Cell growth and colony forming ability were detected with MTT and plate cloning technique.Results: The lentiviral Prdx2 expression vector was successful construc-ted.Overexpression of Prdx2 was verified in SW480 cells with LV-Prdx2 vector.Prdx2 promoted SW480 cell growth and colony forming ability(P<0.05).Conclusion:Ubi-Prdx2-EGFP-Puromycin(LV-Prdx2) vector is successfully constructed,and the SW480/LV-Prdx2 cell line with stable transduction of Prdx2 containing vector is established.Overexpression Prdx2 can significantly promote the proliferation of colorectal cancer SW 480 cells.
2.Construction of lentiviral vector of peroxiredoxin 2 gene mediating RNAi and its effects on the proliferation of SW480 cell
Jihong FENG ; Zhongxue FU ; Kunming WEN ; Weidong LU ; Hao WANG ; Wangsheng CHEN ; Jinbao GUO ; Shouru ZHANG
Chongqing Medicine 2015;(1):14-17,20
Objective To construct a lentiviral expression vector of peroxiredoxin2(PRDX2) RNA interference (RNAi) and to investigate the effect of siRNA of PRDX2 genes on the proliferation of human colonrectal cancer SW480 cell .Methods RNAi tar‐get sequences were designed and synthesized towards the PRDX2 gene sequences .The lentiviral vector pGC‐EGFP‐shPRDX2 was constructed and identified .The vector was transformed into SW480 cells ,and the transfection efficiency was evaluated by fluores‐cence microscopy .The expression of PRDX2 was detected with Quantitative real‐time PCR (qRT‐PCR) and Western blot in the transfected cells .Cell growth and colony forming ability were detected with MTT and plate cloning technique .Results PRDX2 gene lentiviral vector was successfully established and was proved by gene sequencing .The expression of PRDX2 in mRNA and pro‐tein was significantly reduced(P<0 .05) .The PRDX2 mRNA and protein expression in SW480 transfected with lentiviral were sig‐nificantly reduced (P< 0 .05) ,and the ability of growth and proliferation were significantly reduced(P< 0 .05) .Conclusion PRDX2 gene lentiviral vector could be a stable and reliable tool .The proliferation and growth of SW480 cells transfected by pGC‐EGFP‐shPRDX2 could be effectively suppressed ,which could facilitate further investigation of the roles of PRDX2 gene in the de‐velopment and progression of colorectal cancer .
3.The role of α-synuclein ubiquitination in its selectivity of degradation pathway
Zenglin CAI ; Yuanyuan LIU ; Shouru XUE ; Jing XU ; Qingzhi ZHANG ; Xiuming LI
The Journal of Practical Medicine 2015;(11):1758-1762
Objective To investigate SIAH′s role in α-synuclein degradation, formation of Lewy bodies and neuronal death. Methods Proliferative activity of PC12 cells was measures by MTT assay after treatment with MPP and Rapamycin. Western Blot was applied determine the protein expression of LC3-Ⅱ, E1, SIAH-1, P53 and α-synucleinto. PCR was applied to measure protein related mRNA levels. Immunofluorescent techniques were used to detect the distribution of α-synuclein, SIAH-1 and LC3 in cells after SIAH antibody processing. Results MPP+ treatment increased α-synuclein, E1 expression and SIAH-1 activity, however, LC3-Ⅱ, P53 and α-synuclein protein levels decreased significantly. Anti-SIAH-1 antibody treatment reversed this trend, with E1 significantly increased. Rapamycin treatment reduced SIAH-1 and α-synuclein levels in the MPP+ group. SIAH-1 antibody significantly decreased the positive immuno-stain of α-synuclein, SIAH-1 and LC3, suggesting loss of co-localization. Conclusions Anti-SIAH-1 supports the clearance of non-aggregated α-synuclein by the UPS. SIAH plays a key role in the pathogenesis of Parkinson′s disease and is a potential therapeutic target of neurodegenerative diseases.
4.Progress of diagnosis and treatment for gastrointestinal neuroendocrine neoplasms
Miao HE ; Jing FAN ; Ziwei WANG ; Shouru ZHANG
Cancer Research and Clinic 2023;35(4):309-313
Gastrointestinal neuroendocrine neoplasm (GI-NEN) is an important part of digestive system tumor. With the progress of diagnosis and the further understanding of GI-NEN, the diagnostic rate is increasing. However, GI-NEN is a rare tumor, thus clinicians are not familiar with the diagnosis and treatment. Misdiagnosis and missed diagnosis of GI-NEN often occur, and the treatment regimen is not standardized. In recent years, some progresses of the diagnosis and treatment have been made in GI-NEN, especially in the aspects of pathology, imaging diagnosis, radionuclide therapy, targeted therapy and immunotherapy, which have improved the diagnosis rate and efficacy for GI-NEN. This paper reviews recent literatures of GI-NEN, summarizes its epidemiology, pathological diagnosis, image diagnosis, surgery, targeted therapy, radionuclide therapy, chemotherapy, immunotherapy, and separates GI-NEN into metastatic and non-metastatic groups for detailed review to provide a reference for guiding clinical diagnosis and treatment as well as exploring the new treatment regimens of GI-NEN.