1.CD147 monoclonal antibody-mediated nanoparticles for gene therapy to target lung cancer cells
Feilong WU ; Qinglei KONG ; Songwang CAI ; Zhiqiang YE
Chinese Journal of Pathophysiology 2016;32(9):1562-1567
AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells.The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed.METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector.The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope.The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group.Non-transfected cells were used as control group.The cell transfection was carried out with 250 ng plasmids/well in 6-well plate.The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope.The mRNA expression of PKCε was detected by RT-qPCR.The protein expres-sion of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot.The cell proliferation ability was detec-ted with colony formation assay.The cell invasion ability was detected by Transwell method.RESULTS: The expression of
CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining.The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76 ± 0.18)%, (98.51 ±0.32)%, (99.17 ±0.16)% and (99.68 ±0.11)%, respectively.The difference between CP group and control group was statistically significant (P <0.05).No significant difference among CN group, LP group and control group was observed.The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference.The colony number in CP group was significantly smaller than that in control group (P <0.05).The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group.The number of the invading cells in CP group was significantly less than that in control group (P <0.05).The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group.CONCLUSION: Nanogene vector targe-ting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibito-ry effects on the proliferation and invasion of the lung cancer cells.
2.The expression status of ZIC2 is an independent prognostic marker of hepatocellular carcinoma
Kong QINGLEI ; Li WENCHAO ; Hu PAN ; Zeng HUA ; Pan YUHANG ; Zhou TAICHENG ; Hu KUNPENG
Liver Research 2020;4(1):40-46
Background:Zinc finger protein of cerebellum 2(ZIC2)is a transcriptional activator or repressor that is important for the organogenesis of the central nervous system.Previous studies have reported that ZIC2 is widely upregulated in a variety of tumors. Methods:Oncomine database was used to evaluate the expression levels of ZIC2 in hepatocellular car-cinoma(HCC)and normal liver tissues.Quantitative real-time polymerase chain reaction and immu-nohistochemistry were conducted to validate the results from the database.Cox regression analysis and survival curves were performed to assess the survival effect of ZIC2 in HCC. Results:Increased expression of ZIC2 was detected in HCC tissues compared with normal liver tissues.In addition,patients with high ZIC2 levels had a poor prognosis.Multivariate analysis showed that clinical stage(T or M classification)and ZIC2 levels were independent prognostic factors for overall survival.Moreover,a subgroup analysis revealed that patients with moderate or poor grade tumors,T1-2 tumors,N0 tumors,early American Joint Committee on Cancer(AJCC)stage,and old age were more likely to present with overexpression of ZIC2.To conclude,ZIC2 is upregulated in HCC and associated with the histology and survival of HCC patients. Conclusions:The expression status of ZIC2 may serve as an independent prognostic marker of HCC.
3.Effect of decorin on proliferation and collagen type I synthesis of stiff knee joint synovial type B cells
Bangrong XING ; Yuxian CHEN ; Hua ZENG ; Xiaoyan YANG ; Caiqian LIANG ; Weixiong ZHOU ; Qinglei KONG ; Jianhua HAN ; Yongbiao ZHANG
Chinese Journal of Pathophysiology 2015;(7):1277-1281
AIM:To explore the effects of decorin on procollagen type I (PcI), mRNA expression,collagen type I synthesis and proliferation of synovial type B cells of stiff knee joint synovial membrane.METHODS:Type B cells of synovial membrane were isolated from the stiff knee joint synovial membrane and cultured in vitro.The cells were treated with decorin at concentrations of 0.1 mg/L, 5 mg/L and 10 mg/L.After cultured for 24 h, 48 h and 72 h, the cell proli-feration rates were measured by MTT colorimetric determination.Cell cycle distribution and apoptosis were analyzed by flow cytometry.The mRNA level of Pc I was detected by RT-PCR, while collagen type I was measured by Western blot.RE-SULTS:The proliferation of synovial type B cells was significantly inhibited, the percentage of synovial type B cells at G1 phase was significantly increased by 5 mg/L and 10 mg/L decorin (P<0.05), and PcⅠmRNA expression and collagen type I synthesis were significantly decreased.The cells with late apoptosis were not found in control group and experimental groups.CONCLUSION:Recombinant human decorin inhibits synovial type B cell proliferation and decreases PcⅠmRNA expression and collagen type I synthesis in synovial type B cells of stiff knee joint synovial membrane in vitro, suggesting that decorin potentially contributes to the therapy of human knee stiffness.
4.Mechanisms of plasmid-mediated quinolone resistance in Proteus mirabilis from urine
Caiqian LIANG ; Qinglei KONG ; Yongmei FU ; Yaqun FENG ; Xiaoyan YANG
The Journal of Practical Medicine 2017;33(24):4152-4155
Objective To investigate the prevalence of plasmid-mediated quinolone resistance(PMQR) in Proteus mirabilis clinical isolated from urine. Methods Antimicrobial susceptibility test was performed using the microdilution method.And PMQR gene qnrA,qnrB,qnrC,qnrD,qnrS,aac(6′)-Ib-cr,qepA were amplified by PCR,then the PCR positive products were sequenced to identify their genotypes. Results In 41 strains of Proteus mirabilis from urine,the resistant rates to ciprofloxacin and levofloxacin were 41.5% and 29.3%,respec-tively.Among all clinical isolates,qnrA1 gene was detected in 2 strains,qnrB2 gene in 3 strains.PMQR gene qn-rB,qnrC,qnrD,qnrS,aac(6')-Ib-cr and qepA were not detected in all strains.Conclusions Clinical isolates of Proteus mirabilis from urine carry PMQR genes.The prevalent principal genotypes are qnrA1 and qnrB2 in these iso-lates.They are related to low levels resistance toquinolone.
5.Influence and mechanism of c-jun gene silenced by siRNA on liver regeneration in liver cirrhotic rats after hepatectomy
Yuewu YANG ; Xuhui LIU ; Qinglei KONG ; Zhiqiang YE
Chinese Journal of Hepatic Surgery(Electronic Edition) 2015;(3):187-190
ObjectiveTo investigate the influence and mechanism of c-jun gene silenced by siRNA on liver regeneration in liver cirrhotic rats after hepatectomy.MethodsLiver cirrhotic rat models were established and 50 liver cirrhotic rats were randomized into the treatment group and the control group by random number table method with 25 rats in each group. The rats in the treatment group underwent major hepatectomy and were administered with lentivirus carrying plasmid Pcmv6-AC-GFP/ c-jun-siRNA by intraportal injection, while the rats in the control group underwent major hepatectomy and were administered with phosphate buffer saline (PBS) by intraportal injection. The portal pressure, serum ALT and liver weight/body weight of both groups were tested 14 d after surgery. The relative expression of c-jun in the liver and proliferating cell nuclear antigen (PCNA) protein was detected by Western blot. The data between the treatment group and the control group were compared usingt test.ResultsThe average portal pressure of the treatment group after surgery was (1.18±0.11) kPa, which was signiifcantly lower than (1.67±0.24) kPa of the control group (t=-26.74,P<0.05). ALT of the treatment group after surgery was (43±5) U/L, which was signiifcantly lower than (257±14) U/L of the control group (t=-31.11, P<0.05). Liver weight/body weight of the treatment group after surgery was (6.94±0.31)%, which was signiifcantly greater than (2.76±0.14)% of the control group (t=23.57,P<0.05). The relative expression of c-jun and PCNA protein of the treatment group after surgery was respectively 0.143±0.014 and 0.195±0.027, and those of the control group was respectively 0.742±0.057 and 0.029±0.003, where signiifcant difference was observed (t=-37.17, 14.86;P<0.05).ConclusionLentivirus carrying c-jun-siRNA may promote liver regeneration and improve live function in cirrhotic rats after hepatectomy by down-regulating the c-jun in hepatocyte and up-regulating the PCNA protein.
6.Expression of c-Jun in hepatocellular carcinoma and its significance
Yuewu YANG ; Zhiqiang YE ; Qinglei KONG ; Gangjian LUO
Chinese Journal of Hepatic Surgery(Electronic Edition) 2015;(4):250-253
ObjectiveTo investigate the expression of c-Jun in hepatocellular carcinoma (HCC) and its correlation with prognosis.MethodsClinical data of 137 patients with HCC undergoing hepatectomy in the Third Affiliated Hospital of Sun Yat-sen University between Janunary 2010 and December 2013 were retrospectively studied. Among the 137 patients, 112 were males and 25 were females with the age ranging from 26 to 73 years old and the median of 45 years old. The informed consents of all patients were obtained and the local ethical committee approval had been received. The expression of c-Jun in HCC tissues was detected by immunohistochemistry. The patients were divided into the high-expression group (n=75) and low-expression group (n=62) according to the scores of immunohistochemical staining. The clinicopathological parameters and the correlation between c-Jun expression and prognosis of the two groups were analyzed. Comparison on clinicopathological parameters was conducted using Chi-square test and survival analysis was conducted using Kaplan-Meier and Log-rank test.ResultsThe expression of c-Jun in HCC was correlated with HBsAg, alpha-fetoprotein (AFP), tumor size, tumor capsule and tumor vascular invasion (χ2=5.27, 32.68, 4.86, 1.04, 4.62;P<0.05). The 3-, 5-year cumulative survival rate of the high-expression group was respectively 64.14% and 41.20% and those of the low-expression group was respectively 72.04% and 61.21%. The overall survival of the high-expression group was signiifcantly lower than that of the low-expression group (χ2=5.19,P<0.05).Conclusionc-Jun may play an important role in the occurrence and development of HCC and high expression of c-Jun indicates poor prognosis.
7.Adjuvant chemotherapy versus adjuvant concurrent chemoradiotherapy after radical surgery for early-stage cervical cancer: a randomized, non-inferiority, multicenter trial.
Danhui WENG ; Huihua XIONG ; Changkun ZHU ; Xiaoyun WAN ; Yaxia CHEN ; Xinyu WANG ; Youzhong ZHANG ; Jie JIANG ; Xi ZHANG ; Qinglei GAO ; Gang CHEN ; Hui XING ; Changyu WANG ; Kezhen LI ; Yaheng CHEN ; Yuyan MAO ; Dongxiao HU ; Zimin PAN ; Qingqin CHEN ; Baoxia CUI ; Kun SONG ; Cunjian YI ; Guangcai PENG ; Xiaobing HAN ; Ruifang AN ; Liangsheng FAN ; Wei WANG ; Tingchuan XIONG ; Yile CHEN ; Zhenzi TANG ; Lin LI ; Xingsheng YANG ; Xiaodong CHENG ; Weiguo LU ; Hui WANG ; Beihua KONG ; Xing XIE ; Ding MA
Frontiers of Medicine 2023;17(1):93-104
We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.
Female
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Humans
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Uterine Cervical Neoplasms/drug therapy*
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Prospective Studies
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Quality of Life
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Neoplasm Staging
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Chemoradiotherapy
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Chemotherapy, Adjuvant/adverse effects*
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Adjuvants, Immunologic
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Antineoplastic Combined Chemotherapy Protocols/therapeutic use*
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Retrospective Studies