1.Effect of Gold (Au) Nanoparticles Modified by Surface Chemistry on the Proliferation and Migration of Hepatocellular Carcinoma Cells in Vitro.
Jinyong HONG ; Hongmei YIN ; Yang SHEN ; Zhiping YAN ; Jingxia LIU ; Fating ZHOU ; Qing XIA ; Xiaoheng LIU
Journal of Biomedical Engineering 2015;32(2):373-379
Due to the good tumor-targeting and excellent biocompatibility, the drug-loading nanoparticles (NPs) has been widely applied in the diagnosis and treatment of cancer. However, after the NPs are recognized and internalized by cancer cells, the effects of NPs on cell migration behavior were unclear. In the present study, the self-assembly techniques (SAMs) was used to modify gold (Au) nanoparticles (Au NPs) with different chemical functional groups (CH3, OH, COOH and NH2) as model NPs. The dispersion of these groups in solution and the distribution in cells were studied by transmission electron microscope (TEM), respectively, and the proliferation was examined by MTT assay in vitro. The wound-healing and the Transwell assay were used to examine the effect of internalized Au-NPs on HepG2 cells migration. The results showed that different Au-NPs mainly distributed at the edge of the vesicle membrane and the gap between cells. The Au-NPs resulted in decreased cell viability in a concentration-depended manner. In addition, the results of wound-healing and Transwells assay indicated that the internalization of the NH2-NPs and OH-NPs would inhibit cell migration compared with those in the control group.
Carcinoma, Hepatocellular
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metabolism
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Cell Movement
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Cell Proliferation
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Cell Survival
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Gold
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Hep G2 Cells
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Humans
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Liver Neoplasms
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metabolism
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Metal Nanoparticles
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chemistry
2.Effect of fluid shear stress on the cellular morphology and tight junction of laryngeal squamous carcinoma Hep2 cells.
Fating ZHOU ; Hongmei YIN ; Shuangfeng LIU ; Yang SHEN ; Jinyong HONG ; Qing XIA ; Xiaocheng LIU
Journal of Biomedical Engineering 2015;32(1):104-109
This paper is aimed to investigate the effect of fluid shear stress on the tight junction of laryngeal squamous carcinoma (Hep2) cells and to explore the potential molecular mechanism. Hep2 cells were selected and subjected to the fluid shear stress of 1.4 dyn/cm2 for different time, respectively. The morphological changes of Hep2 cells under shear stress were observed using inverted microscope. The cell-cell junctions were examined by transmission electron microscope (TEM). The expressions of tight junction proteins (including Occludin, Claudin-5 and ZO-1) and the distribution of Claudin-5 were examined by Western blot assay and laser scanning confocal microscope, respectively. The results indicated that Hep2 cells turned to spindle-like shapes after exposed to shear stress, and showed the trend of the recovering to original shapes when the shear stress was cancelled. The cell-cell junctions were tight under the shear flow condition, and the permeability was reduced under the condition of 1.4 dyn/cm shear flow. The expressions of tight junction proteins were enhanced with increased duration of shear flow, but reduced after removing shear flow. The result of Claudin-5 expression by immufluorescence assay was consistent with that by Western blot. The Claudin-5 mainly distributed in the cytoplasm under static condition, while it located at the intercellular after shear flow stimulation, and it appeared intercellular and cytoplasm after stopping shear flow stimulation. Therefore, it can be concluded that shear stress changes the morphology of laryngeal squamous carcinoma Hep2 cells, and upregulates the tight junction.
Blotting, Western
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Carcinoma, Squamous Cell
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pathology
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Claudin-5
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metabolism
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Hep G2 Cells
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Humans
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Laryngeal Neoplasms
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pathology
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Occludin
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metabolism
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Stress, Mechanical
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Tight Junctions
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Zonula Occludens-1 Protein
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metabolism
3.Influence of Tumor Microenvironment of Hepatocellular Carcinoma on the Proliferation of Vascular Endothelial Cells and Vascular Angiogenesis Ability.
Qin XIA ; Hongmei YIN ; Yang SHEN ; Jingxia LIU ; Zhiping YAN ; Jinyong HONG ; Fating ZHOU ; Xiaoheng LIU
Journal of Biomedical Engineering 2015;32(3):612-617
To study the potential molecular mechanism of tumor angiogenesis in its microenvironment, we investigated the effects of HepG2 conditioned medium on the proliferation of vascular endothelial cell and vascular angiogenesis in our laboratory. Human umbilical vein endothelial EA. hy926 cells were co-cultured with HepG2 conditioned medium in vitro. The proliferation and the tubulogenesis of EA. hy926 cells were detected by teramethylazo salt azole (MTT) and tube formation assay, respectively. The results showed that the survival rate of the EA. hy926 cells was significantly increased under the co-culture condition. HepG2 conditioned medium also enhanced the angiogenesis ability of EA. hy926 cells. In addition, the expressions of intracellular VEGF and extracellular VEGFR (Flk-1) were regulated upward in a time-dependent manner. In conclusion, the proliferation of vascular endothelial cells and Vascula angiogenesis were improved under the condition of indirect co-culture.
Carcinoma, Hepatocellular
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pathology
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Cell Proliferation
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Coculture Techniques
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Culture Media, Conditioned
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Endothelial Cells
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cytology
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Hep G2 Cells
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Human Umbilical Vein Endothelial Cells
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Humans
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Liver Neoplasms
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pathology
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Neovascularization, Pathologic
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Tumor Microenvironment
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Vascular Endothelial Growth Factor A
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metabolism
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Vascular Endothelial Growth Factor Receptor-2
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metabolism
4.Integrins mediate the migration of HepG2 cells induced by IL-8
Yi WANG ; Zhenhua YANG ; Xuedan LI ; Honggen QIU ; Jun XU ; Fating ZHOU
Military Medical Sciences 2016;40(8):623-627
[ Abstract] Objective To investigate the effect of IL-8 on the viability and migration of HepG2 cells and the potential effect of integrins (αsubunits ) on the migration of HepG2 cells.Methods HepG2 cells were stimulated by IL-8 at different concentrations ranging from 0 to 125 ng/ml in vitro.MTT assay was preformed to detect the viability of HepG2 cells.Scratch wound migration assay was used to explore the effect of IL-8 on the migration of HepG2 cells at the time points of 0, 4, 8, 12 and 24 h,respectively.Transwell assay was conducted to detect the vertical migration of HepG2 cells after stimulation with IL-8 for 24 h.The F-actin of HepG2 cells was observed by immunofluorescence analysis after treatment with IL-8 at different concentrations.The expression of αsubunits of integrins in HepG2 cells treated with IL-8 for 8h was detected using Western boltting assays.Results Compared with the control group, HepG2 cells treated with IL-8 showed improved proliferation activities, but there was no significant difference between the groups of HepG2 cells treated with IL-8 at different concentrations.The cell scratch healing assays and Transwell analysis both indicated that IL-8 promoted the migration of HepG2 cells in a dose-dependent manner.Meanwhile, the cytoskeleton of HepG2 cells was rearranged and the number of filopodium-like protrusions (FLPs) was increased rapidly after treatment with IL-8.Western blotting analysis showed that IL-8 up-regulated the expression of αsubunit of integrins and there was a concentration difference betweenαsubunits.Conclusion IL-8 promotes the migration of HepG2 cells by up-regulating the expression ofαsubunits, and different αsubunits may play different roles.
5.ThevalueofMRIIDEALIQtechniquetoevaluate myelosuppressionduringradiotherapyandchemotherapyincervicalcancer
Jingling LI ; Yunfeng MU ; Shun QI ; Haitao ZHAO ; Jianmin ZHENG ; Fang REN ; Fating YANG
Journal of Practical Radiology 2019;35(4):643-646
Objective ToexplorethevalueofMRIiterativedecompositionofwaterandfatwithechoasymmetryandleastsquares estimationquantificationsequence(IDEALIQ)techniquetoevaluatemyelosuppressionduringradiotherapyandchemotherapyincervicalcancer. Methods 25femalesubjectswereenrolledinthisstudy,whowereclinicallydiagnosedascervicalcancerandacceptedtheradiotherapyand chemotherapy.AllthesubjectswereperformedwithsagittalMRIIDEALIQscansineachweek’streatmentandattheendofwhole fiveweeks’therapy,soeachpatienthad6timesMRIscans.ROIweremanuallyplacedonL4,L5andS1vertebralbodyandsubcutaneousfatto measurethefatfraction.ThefatfractioncolorimageswerereconstructedonaAW (AdvantageWorkstation)4.6workstation.Results Asthe radiationandchemotherapyprocess,thevaluesoffatfractionincreasedprogressivelyonL4,L5andS1vertebralbody(P<0.001), whilethefatfractionvaluesinsubcutaneousfatappearedstableallthetime(P=0.987).Conclusion MRIIDEALIQtechniquecan evaluatethereal-timefatfraction,andradiotherapyandchemotherapyplansmaybeoptimizedaccordingtothefatfractionresult.
6.Clinical analysis of cervical lesions in 1 337 postmenopausal women diagnosed during screening
Weihong QI ; Qin LI ; Xiaoke YANG ; Linlin MA ; Yong LI ; Fating HUANG ; Bing QU ; Jianxin PU
Chinese Journal of Geriatrics 2023;42(7):804-809
Objective:To screen and diagnose cervical lesions in middle-aged and elderly postmenopausal women and analyze the characteristics of cervical lesions in middle-aged and elderly women.Methods:A retrospective analysis was conducted on the screening data of 1 337 postmenopausal women aged between 50 and 86 years who visited the outpatient clinics of the gynecology department of Beijing Hospital between January, 2015 and December, 2019.Participants were divided into groups based on age, with 787 in the 50-59 years old group, 483 in the 60-69 years old group, 60 in the 70-79 years old group, and 7 in the 80-89 years old group.Exfoliated cervical cells were collected, and high-risk human papillomavirus(HPV)detection and liquid-based cervical cytology examination were performed to confirm the diagnosis based on the pathological results of cervical biopsy.Cervical lesions were analyzed.Results:53.0%(708)of the 1 337 enrolled patients made their hospital visits because of symptoms.There were 666 patients(49.8%)with cervical lesions, including 357 with low-grade squamous intraepithelial lesions(LSIL), 252 with high-grade squamous intraepithelial lesions(HSIL)and 57(4.3%)with cervical cancer.The 70-79 group showed the highest incidence of HSIL at 30.0%(18/60), while the 50~59 group had the highest incidence of LSIL at 30.6%(241/787), and there were significant differences in different types of cervical lesions between different age groups( χ2=28.919, P<0.01). Liquid-based cervical cytology examination revealed a positive rate of 51.6%(166/322)for LSIL, a positive rate of 70.8%(143/202)for HSIL, and a rate of 71.4%(25/35)for cervical cancer.There were significant differences in LSIL, HSIL and other above-mentioned positive rates of lesions( P<0.05). In the cervical malignancy group, cytological high-grade squamous epithelial lesions made up 51.4%(18/35). Intraepithelial lesions or cancerous lesions represented 29.2%(59/202)in the HSIL group.The rate of intraepithelial lesions or cancerous lesions in the LSIL group was 48.4%(156/322), with statistically significance in these differences( χ2=118.806, P<0.01). Chronic cervicitis was found in 52.9%(109)of 206 patients with atypical squamous cells and positive HPV, and in 76.2%(16)of 21 patients without HPV.HPV examination was completed in 1 301 patients, with 69.4%(903)having infections by one HPV strain and 30.6%(398)having infections by mixed HPV strains.Biopsy results showed that the rates of HPV infections in LSIL, HSIL and cancer patients were 84.6%(302/357), 95.9(185/193)and 100%(54/54), respectively, and the differences in these pathological results were statistically significant( P<0.01). HPV16 was the most common subtype in single HPV strain infections.The most common subtypes of HPV infections in cervical lesions with HSIL and above were HPV16 and HPV18.The agreement rate was 52.4%(187/357)for LSIL, 60.7%(153/252)for HSIL, and 64.9%(37/57)for cervical cancer between diagnoses based on cervical biopsy and those based on colposcopy.As the severity of lesions increased, the accuracy of colposcopy also increased.Differences between colposcopic diagnosis and pathological diagnosis on different types of cervical lesions were statistically significant( χ2=550.382, P<0.01).1 310 patients underwent endocervical curettage, identifying 120 cases of HSIL and 39 cases of cervical cancer, with a detection rate of 55.8%(159/285). Conclusions:The incidence of cervical cancer is high in elderly women.The incidences of high-grade lesions and cervical cancer in patients with atypical squamous cells plus HPV infections in the cervix are also high.Postmenopausal patients with cervical lesions are mainly infected by a single HPV subtype, with subtype 16 being the most common one.With increasing severity of lesions, the accuracy of colposcopy-based diagnosis increases and the rate of missed diagnosis decreases.Cervical curettage is an important means to reduce missed diagnosis with colposcopy.