Induction of apoptosis and G2/M cell cycle arrest by oridonin in human gastric cancer BGC-823 cells

Aim To investigate in vitro apoptosis-induction effects of oridonin on gastric tumor cells BGC-823 and its effects on cell cycle, mitochondrial membrane potential and intracellular Ca^2＋ to shed light on the mode of its anticancer action. Methods The MTT method was used to investigate the inhibitory effect of oridonin on BGC-823 cells. The apoptosis-induction effect was evaluated by confocal laser microscopy and flow cytometry. The change of mitochondrial membrane potential and the increase of intracellular Ca^2＋ were assessed by fluorescence probe rhodamine123 and Fluo 3-AM, respectively, with flow cytometry. The expression of apoptosis and cell cycle related proteins was studied using western blotting. Results Oridonin inhibited BGC-823 cells growth with IC50 of 22.21 p, mol.L^-1. It induced apoptosis in a dose-dependent manner. In addition, it decreased mitochondria membrane potential, increased intracellular Ca^2＋, and activated pro-caspase 3. BGC-823 cells were arrested in G2/M cell cycle phase with lower expression of cyclin A protein. The up-regulation of p53 was observed before apoptosis and cell cycle arrest occurred. Conclusion Oridonin inhibits the proliferation of BGC-823 cells through G2/M cell cycle arrest and apoptosis induction, which is mediated by influx of Ca^2＋, up-regulation of p53, activation of caspase-3, and down-regulation of cyclin A.

Apoptosis mechanisms of human gastric cancer cell line MKN-45 infected with human mutant p27

AIM: To explore the inducing effect of human mutant p27 gene on the apoptosis of the human gastric cancer cell line MKN-45 and its associated mechanisms. METHODS: The recombinant adenovirus Ad-p27mt was constructed to infect the human gastric cancer cell line MKN-45. Using flow cytometry, TUNEL assay and DNA fragment analysis, we measured the apoptotic effect of Ad-p27mt on the human gastric cancer cells. RESULTS: Ad-p27mt was successfully constructed and the infection efficiency reached 100%. After 18 h of infection, we observed an apoptotic hypodiploid peak on the flow cytometer before G1-S and apoptotic characteristic bands in the DNA electrophoresis. The apoptotic rate detected by TUNEL method was significantly higher in the Ad-p27mt group (89.4±3.12%)compared to the control group (3.12±0.13%, P ＜ 0.01).CONCLUSION: Human mutant p27 can induce apoptosis of the human gastric cancer cells in vitro.

Human induced pluripotent stem cells labeled with fluorescent magnetic nanoparticles for targeted imaging and hyperthermia therapy for gastric cancer

Objective: Human induced pluripotent stem(i PS) cells exhibit great potential for generating functional human cells for medical therapies. In this paper, we report for use of human i PS cells labeled with fluorescent magnetic nanoparticles(FMNPs) for targeted imaging and synergistic therapy of gastric cancer cells in vivo. Methods: Human i PS cells were prepared and cultured for 72 h. The culture medium was collected, and then was coincubated with MGC803 cells. Cell viability was analyzed by the MTT method. FMNP-labeled human i PS cells were prepared and injected into gastric cancer-bearing nude mice. The mouse model was observed using a small-animal imaging system. The nude mice were irradiated under an external alternating magnetic field and evaluated using an infrared thermal mapping instrument. Tumor sizes were measured weekly. Results: iP S cells and the collected culture medium inhibited the growth of MGC803 cells. FMNP-labeled human iP S cells targeted and imaged gastric cancer cells in vivo, as well as inhibited cancer growth in vivo through the external magnetic field. Conclusion: FMNP-labeled human i PS cells exhibit considerable potential in applications such as targeted dual-mode imaging and synergistic therapy for early gastric cancer.

Human epidermal growth factor receptor 2 expression level and combined positive score can evaluate efficacy of advanced gastric cancer

BACKGROUND Although treatment options for gastric cancer(GC)continue to advance,the overall prognosis for patients with GC remains poor.At present,the predictors of treatment efficacy remain controversial except for high microsatellite instability.AIM To develop methods to identify groups of patients with GC who would benefit the most from receiving the combination of a programmed cell death protein 1(PD-1)inhibitor and chemotherapy.METHODS We acquired data from 63 patients with human epidermal growth factor receptor 2(HER2)-negative GC with a histological diagnosis of GC at the Cancer Hospital,Chinese Academy of Medical Sciences between November 2020 and October 2022.All of the patients screened received a PD-1 inhibitor combined with chemotherapy as the first-line treatment.RESULTS As of July 1,2023,the objective response rate was 61.9%,and the disease control rate was 96.8%.The median progression-free survival(mPFS)for all patients was 6.3 months.The median overall survival was not achieved.Survival analysis showed that patients with a combined positive score(CPS)≥1 exhibited an extended trend in progression-free survival(PFS)when compared to patients with a CPS of 0 after receiving a PD-1 inhibitor combined with oxaliplatin and tegafur as the first-line treatment.PFS exhibited a trend for prolongation as the expression level of HER2 increased.Based on PFS,we divided patients into two groups:A treatment group with excellent efficacy and a treatment group with poor efficacy.The mPFS of the excellent efficacy group was 8 months,with a mPFS of 9.1 months after excluding a cohort of patients who received interrupted therapy due to surgery.The mPFS was 4.5 months in patients in the group with poor efficacy who did not receive surgery.Using good/poor efficacy as the endpoint of our study,univariate analysis revealed that both CPS score(P=0.004)and HER2 expression level(P=0.015)were both factors that exerted significant influence on the efficacy of treatment the combination of a PD-1 inhibitor and chemotherapy in patients with advanced GC(AGC).Finally,multivariate analysis confirmed that CPS score was a significant influencing factor.CONCLUSION CPS score and HER2 expression both impacted the efficacy of immunotherapy combined with chemotherapy in AGC patients who were non-positive for HER2.

Anticancer effect of Jinlongshe granules on in situ-transplanted human MKN-45 gastric cancer in nude mice and xenografted sarcoma 180 in Kunming mice and its mechanism

AIM： To study the antitumor effect of Chinese compound Jinlongshe （JLS） granules on sarcoma 180 and MKN-45 human gastric cancer cell lines in vivo and its mechanism. METHODS： After establishment of S180 sarcoma （S180） and MKN-45 gastric cancer model of nude mice, the tumor-bearing mice were divided into 5 groups at random. Three experimental groups were respectively given the aqueous extract of JLS granules at doses of 120 g, 60 g and 20 g/（kg per 6/wk,i.g） for 3 wk in S180 and 6 wk in nude mice model. Positive control was given cyclophosphamide （Cy） at a dose of 50 mg/（kg per 3/wk, i.g） for 3 wk in S180 models and 5-Fluorouracil （5-FU） 20 mg/（kg per 3/wk, i.g） for 3 wk in nude mice model. Negative control was given normal saline （NS） at a dose of 0.18 g/（kg per 6/wk, i.g） respectively. After 3 wk in mice bearing S180 tumor and 6 wk in nude mice model, the experimental animals were sacrificed and the masses of tumor were weighed, and the rates of tumor inhibition of each treated group were calculated respectively. To determine the antitumor mechanisms, the morphological changes, cell cycle and apoptosis were observed in MKN-45 nude mice model. Annexin V-FITC/PI double staining FCM assay was used to further determine the live cells, apoptotic cells, necrotic cells and debris. RESULTS： The inhibitory rates of JLS granules at the doses of 20 g/kg, 60 g/kg and 120 g/kg were 50.31%, 55.94% and 68.13% （P 〈 0.01） in nude mice models and 40.90%, 50.32% and 58.46% （P 〈 0.01） in S180 model. The inhibitory rate of Cy was 85.22% in S180 models and the inhibitory rate of 5-FU was 53.43% in nude mice model （P 〈 0.01）. Nuclear chromatin and margination were observed under a transmission electron microscope （TEM）. The G0/G1 phase was arrested, typical apoptotic peak appeared, the apoptotic rate was 22.81%-38.54% in three JLS granule-treated groups. Annexin V-FITC/PI double staining FCM assay showed that the apoptotic cells were 4.36%, 3.08% and 7.08% in three dosages, most cells were localized in the low right quadrant. CONCLUSION：Jinlongshe granules possess anti-tumor effects on experimental tumor models in vivo, and apoptosis induction is one of its anti-tumor mechanisms.

胃癌MKN-28肿瘤细胞系SP细胞亚群初步分析

目的:分析胃癌细胞株MKN-28中是否包含肿瘤干细胞相关的SP(side population)细胞亚群.方法:采用免疫荧光方法检测干细胞标记ABcG2在胃癌细胞株MKN-28的表达;制备MKN-28细胞悬液,经Hoechst33342染色,流式细胞仪分析SP亚群.结果:ABCG2在胃癌细胞系MKN-28中有少量表达,ABCG2阳性细胞约占1.7%.SP亚群占MKN-28细胞的0.25%,维拉帕米阻断后减少为0.05%.结论:人胃癌细胞株MKN-28中存在SP细胞亚群,提示胃癌干细胞的存在.

三联体基序包含蛋白28在胃癌组织中的表达及其对胃癌HGC-27细胞侵袭、迁移、增殖和硼替佐米诱导的细胞凋亡的影响

目的:探讨三联体基序包含蛋白28(TRIM28)在胃癌组织中的表达及其对胃癌HGC-27细胞侵袭、迁移、增殖和硼替佐米(BTZ)诱导的细胞凋亡的影响。方法:收集10例胃癌患者的癌组织和癌旁组织,采用免疫组化和实时荧光定量PCR(RT-qPCR)分别检测TRIM28阳性率及mRNA的表达水平。采用RNA干扰技术敲低人胃癌细胞株HGC-27中的TRIM28表达,采用Transwell和划痕愈合实验探究敲低TRIM28表达对胃癌细胞的侵袭、迁移能力的影响,采用MTT法检测细胞增殖能力,采用流式细胞术检测细胞的凋亡情况。结果:胃癌组织中TRIM28阳性率及其mRNA表达水平高于癌旁组织(均P<0.05)。敲低TRIM28表达可抑制胃癌细胞的侵袭和迁移能力。敲低TRIM28表达显著抑制了胃癌细胞的增殖能力,并显著增强了BTZ诱导的细胞凋亡。结论:TRIM28在胃癌组织过表达,且与胃癌HGC-27细胞的侵袭、迁移、增殖和凋亡有关,同时影响BTZ的抗肿瘤作用。

基于网络药理学的川楝素调控MAPK通路诱导人胃癌MKN-28细胞凋亡

目的:基于网络药理学探索川楝素对胃癌MKN-28细胞增殖的影响及其分子机制。方法:通过网络药理学筛选出川楝素治疗胃癌的分子通路;MTT法观察川楝素对人胃癌MKN-28细胞增殖抑制的量效关系;Hoechst 33258染色法观察川楝素对细胞凋亡的影响;WesternBlot法检测凋亡及丝裂原活化蛋白激酶(mitogen-activatedproteinkinase,MAPK)信号通路相关蛋白表达。结果:网络药理学方法提示川楝素通过MAPK信号通路作用于胃癌,川楝素能显著抑制人胃癌MKN-28细胞增殖,并能诱导人胃癌MKN-28细胞凋亡。WesternBlot实验提示,随着给药剂量增加,CleavedCaspase-3、Cleaved-PARP、p38和p-p38表达量递增。结论:川楝素能高效抑制人胃癌MKN-28细胞增殖;川楝素是通过激活MAPK通路,诱导细胞凋亡,发挥其抗肿瘤药理作用。

Telomerase and hTERT: Can they serve as markers for gastric cancer diagnosis?

AIM: To investigate telomerase activity and human telomerase reverse transcriptase (hTERT) expression in normal human gastric mucosal epithelial cells (nhGMECs) and fibroblasts (nhGMFs).

Expression of annexin II in gastric carcinoma and its role in gastric cancer metastasis

AIM To investigate the expression of annexin II in gastric carcinoma and its role in the metastasis of gastric cancer.METHODS The expression of annexin II in 51 cases of gastric carcinoma and 24 cases of adjacent tissues was detected by immunohistochemistry. The relationship between annexin II and clinical features of gastric cancer was analyzed. Annexin II specific si RNA was used to inhibit the expression of annexin II in gastric cancer HGC-27 cells, and the effects of annexin II on the migration and secretion of matrix metalloproteinases(MMPs) were observed.RESULTS The positive rate of annexin II protein was 82.4% in gastric cancer tissues and 37.5% in adjacent tissues. There was significant difference between the two groups(P < 0.01); and the positive expression of annexin II was not related to the sex and age of the patients(P > 0.05). The expression of annexin IIprotein was correlated with tumor size, histological differentiation, TNM stage, Lymph node metastasis and other clinical features were significantly correlated, the difference was statistically significant(P < 0.05). Inhibition of annexin II expression, gastric cancer HGC-27 cells migration and secretion of MMPs were significantly decreased, the difference was statistically significant(P < 0.05).CONCLUSION Annexin II is highly expressed in gastric cancer tissues, annexin II protein expression is related to tumor size, histological differentiation, TNM staging, lymph node metastasis and other clinical features were significantly correlated. Annexin II high expression can promote the invasion and metastasis of gastric cancer.

Selective inhibition of cell growth by activin in SNU-16 cells

AIM： To investigate whether activin regulates the cell proliferation of human gastric cancer cell line SNU-16 through the mRNA changes in activin receptors, Smads and p21^CIP1/WAF1. METHODS： The human gastric cancer cell lines were cultured, RNAs were purified, and RT-PCRs were carried out with specifically designed primer for each gene. Among them, the two cell lines SNU-5 and SNU-16 were cultured with activin A for 24, 48 and 72 h. The cell proliferation was measured by MTT assay. For SNU-16, changes in ActRIA, ActRIB, ActRIIA, ActRIIB, Smad2, Smad4, Smad7, and p21^CIP1/WAF1 mRNAs were detected with RT-PCR after the cells were cultured with activin A for 24, 48 and 72 h. RESULTS： The proliferation of SNU-16 cells was down regulated by activin A whereas other cells showed no change. Basal level of inhibin/activin subunits, activin receptors, Smads, and p21^CIP1/WAF1 except for activin βB mRNAs was observed to have differential expression patterns in the human gastric cancer cell lines, AGS, KATO III, SNU-1, SNU-5, SNU-16, SNU-484, SNU-601, SNU-638, SNU-668, and SNU-719. Interestingly, significantly higher expressions of ActR IIA and IIB mRNAs were observed in SNU-16 cells when compared to other cells. After activin treatment, ActR IA, IB, and IIA mRNA levels were decreased whereas ActR IIB mRNA level increased in SNU-16 cells. Smad4 mRNA increased for up to 48 h whereas Smad7 mRNA increased sharply at 24 h and returned to the initial level at 48 h in SNU-16 cells. In addition, expression of the p21^CIP1/WAF1 the mitotic inhibitor, peaked at 72 h after activin treatment in SNU-16 cells. CONCLUSION： Our results suggest that inhibition of cell growth by activin is regulated by the negative feedback effect of Smad7 on the activin signaling pathway, and is mediated through p21^CIP1/WAF1 activation in SNU-16 cells.

转录因子Snail对胃癌MKN-28细胞的增殖、凋亡及侵袭的影响

目的:探讨转录因子Snail对胃癌MKN-28细胞的影响。方法:取胃癌MNK-28细胞株的Snail基因进行沉默研究,以顺铂对MNK-28细胞进行处理,并选用CCK-8法不同刺激因素组细胞的增殖情况进行检测,同时选用透射电镜对各组细胞凋亡情况进行观察,而各组细胞侵袭能力则以Transwell小室进行观察。结果:(1)细胞增殖情况分析,顺铂处理及慢病毒沉默Snail基因均可见细胞增殖抑制现象,而shRNA+Snail+顺铂组细胞增殖抑制率显著高于其他组(p <0.05);(2)细胞凋亡情况,与单纯MKN-28细胞组比较,不同刺激因素组细胞凋亡程度均显著升高(p <0.05);(3)细胞侵袭实验分析,shRNA+Snail+顺铂组细胞迁出的细胞数较其他组明显降低(p <0.05)。结论:转录因子Snail可对胃癌MKN-28细胞的增殖情况产生影响,沉默Snail基因有利于加速细胞凋亡,提高化疗药敏感性,可在临床上推广。

Effects of human microRNA-181a-5p on proliferation and migration of gastric cancer cells

Objective To preliminarily explore the effects of human microRNA-181a on migration of gastric cancer cells and its mechanism.Methods The expression of miRNA-181a-5p in gastric cancer cell line GC9811 and peritoneal high metastasis gastric cancer cell line GC9811-P were tested by quantitative real-time polymerase chain reaction(qRT-PCR).GC9811 cell line was

金龙蛇颗粒对裸鼠原位移植人胃癌MKN-45细胞凋亡的影响

目的：评价金龙蛇颗粒对裸鼠原位移植入胃癌MKN-45细胞的抑制作用.方法：50只裸鼠建立原位移植人胃癌MKN-45细胞模型,随机分为模型组,金龙蛇颗粒高、中、低剂量组和5-FU干预组.各组裸鼠予以相应药物实施干预.观察各组荷瘤裸鼠一般情况,肿瘤生长情况及抑瘤率.流式细胞仪检测肿瘤细胞周期分布及细胞凋亡情况.采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-fluorescein isothiocyanate/propidium iodide,Annexin V-FITC/PI）双标记染色法鉴别早期凋亡细胞、晚期凋亡细胞和坏死细胞.电镜下观察肿瘤细胞的超微结构变化.结果：金龙蛇颗粒高、中、低剂量组的抑瘤率分别为68.13%、55.94%和50.31%,呈剂量依赖效应,与5-FU干预组抑瘤率比较,差异无统计学意义.金龙蛇颗粒各剂量组肿瘤细胞凋亡率为22.81%～38.54%,亦呈剂量依赖效应,且肿瘤细胞主要被阻滞于G0/G1期.AnnexinV-FITC/PI双标记染色结果显示,金龙蛇颗粒各剂量组以早期凋亡细胞居多,5-FU干预组则以晚期凋亡细胞和坏死细胞居多.结论：金龙蛇颗粒对裸鼠原位移植人胃癌MKN-45细胞具有较好的抑制作用,促进MKN-45细胞凋亡可能是其抗肿瘤治疗的主要作用机制之一.

槐耳醇提物抑制人胃癌细胞MKN-45增殖的实验研究

目的观察槐耳不同提取部位对人胃癌MKN-45细胞增殖的影响,并筛选其有效部位。方法用系统溶剂法将槐耳提取为石油醚、乙酸乙酯、正丁醇、乙醇、水五个部位后,用CCK-8显色法,检测不同部位不同浓度的槐耳醇提取物对MKN-45增殖的作用。结果槐耳石油醚、乙酸乙酯、正丁醇、乙醇部位为抗肿瘤有效部位,其中以正丁醇部位效果最佳,其对MKN-45细胞半数抑制率(IC50)为56.142μg·m L-1。结论槐耳醇提物能抑制人胃癌MKN-45细胞增殖,其最有效部位为槐耳正丁醇部位。

MAPK通路在红托竹荪多糖诱导人胃癌MKN-45细胞凋亡中的作用机制研究

目的:研究红托竹荪多糖对人胃癌MKN-45细胞增殖与凋亡的影响,并基于丝裂原激活的蛋白激酶(MAPK)信号通路探讨其可能的作用机制。方法:体外培养MKN-45细胞,分为对照组,红托竹荪多糖低、中及高剂量组,采用终浓度分别为0、4、6及8 mg/mL的红托竹荪多糖作用于细胞48 h后,采用细胞计数法检测红托竹荪多糖对人胃癌MKN-45细胞增殖的影响;采用倒置显微镜观察不同浓度的红托竹荪多糖作用于人胃癌MKN-45细胞48 h后细胞形态学的变化。设立实验组的红托竹荪多糖终浓度为8 mg/mL,采用膜联蛋白V-异硫氰酸荧光素/碘化丙啶双染色流式细胞技术检测细胞的凋亡率;采用流式细胞术检测细胞周期分布;采用蛋白质印迹法检测红托竹荪多糖作用胃癌细胞后p38蛋白激酶(p38)、磷酸化p38蛋白激酶(p-p38)、细胞外调节蛋白激酶(ERK)、磷酸化细胞外调节蛋白激酶(p-ERK)、C-JUN氨基末端激酶(JNK)、磷酸化C-JUN氨基末端激酶(p-JNK)、B淋巴细胞瘤-2蛋白(Bcl-2)和活化的半胱氨酸蛋白酶-3(cleaved Caspase-3)蛋白的表达,即与3-磷酸甘油醛脱氢酶(GADPH)灰度值比值。结果:作用48 h后,不同浓度的红托竹荪多糖(4、6及8 mg/mL)对人胃癌MKN-45细胞增殖的抑制率分别为(8.27±5.99)%、(21.83±5.77)%和(56.95±5.21)%;在检测的浓度范围下,随着浓度的升高,红托竹荪多糖对MKN-45细胞增殖的抑制率逐渐升高。倒置显微镜下显示,经8 mg/mL的红托竹荪多糖作用48 h后,MKN-45细胞密度与对照组相比明显减少。流式细胞术检测结果显示,与对照组相比,实验组人胃癌MKN-45细胞凋亡率显著升高(P<0.05);细胞周期结果显示,实验组G_(1)期细胞比例明显减少(P<0.05),G_(2)期细胞比例明显增多(P<0.05),S期细胞比例明显减少(P<0.05);蛋白质印迹法结果显示,与对照组相比,实验组MKN-45细胞中cleaved Caspase-3的表达有升高趋势(P<0.05),Bcl-2的表达有降低趋势(P<0.05),p-P38/P38比值、p-ERK/ERK比值与p-JNK/JNK比值均低于对照组(P<0.05),上述差异均有统计学意义。结论:红托竹荪多糖在体外能抑制人胃癌MKN-45细胞增殖并诱导细胞凋亡,使肿瘤细胞阻滞在G_(2)/M期,其作用可能通过调控MAPK信号通路实现。

槐耳清膏抑制小鼠模型人胃癌MKN-45细胞生长和转移的实验研究

目的研究槐耳清膏对人胃癌MKN-45细胞在裸鼠体内生长和转移的抑制作用,并探讨其作用机制。方法采用BALB/c裸鼠建立人胃癌MKN-45细胞淋巴结转移模型,随机分为槐耳清膏组、空白对照组,每组20只。建模后,每5天测量裸鼠荷瘤并计算肿瘤相对体积(RTV);用药5周后处死裸鼠,Real-time q PCR检测肿瘤组织Wnt、β-catenin、E-cadherin、N-cadherin和VEGF表达水平。结果槐耳清膏组末次测得的足垫瘤RTV为(662.75±450.66)mm3,明显小于空白对照组的(1611.85±611.83)mm3(P<0.01)。槐耳清膏组淋巴结转移率8/20(40%)与对照组15/20(70%)相比,差异有统计学意义(P<0.05)。槐耳清膏组肿瘤组织Wnt、β-catenin、N-cadherin和VEGF的△CT值分别为(11.36±1.32)、(11.31±1.76)、(12.82±2.22)、(13.22±1.52),均高于对照组的(9.48±2.07)、(9.37±1.88)、(7.74±2.64)、(11.04±1.61),而E-cadherin的△CT值(8.26±2.39)低于对照组的(10.64±1.48)(P<0.05)。结论槐耳清膏可抑制人胃癌MKN-45细胞在裸鼠上的生长及转移,其机制可能与抗血管生成和阻碍上皮间质转化(EMT)过程有关。

A novel animal model for in vivo study of liver cancer metastasis

AIM:To establish an animal model with human hepatocyte-repopulated liver for the study of liver cancer metastasis.METHODS:Cell transplantation into mouse livers was conducted using alpha-fetoprotein(AFP)-producing hu-man gastric cancer cells(h-GCCs) and h-hepatocytes as donor cells in a transgenic mouse line expressing urokinase-type plasminogen activator(uPA) driven by the albumin enhancer/promoter crossed with a severe combined immunodeficient(SCID) mouse line(uPA/SCID mice).Host mice were divided into two groups(A and B).Group A mice were transplanted with h-GCCs alone,and group B mice were transplanted with h-GCCs and h-hepatocytes together.The replacement index(RI),which is the ratio of transplanted h-GCCs and h-hepatocytes that occupy the examined area of a histological section,was estimated by measuring h-AFP and h-albumin concentrations in sera,respectively,as well as by immunohistochemical analyses of h-AFP and human cytokeratin 18 in histological sections.RESULTS:The h-GCCs successfully engrafted,repopulated,and colonized the livers of mice in group A(RI = 22.0% ± 2.6%).These mice had moderately differentiated adenocarcinomatous lesions with disrupted glandular structures,which is a characteristics feature of gastric cancers.The serum h-AFP level reached 211.0 ± 142.2 g/mL(range,7.1-324.2 g/mL).In group B mice,the h-GCCs and h-hepatocytes independently engrafted,repopulated the host liver,and developed colonies(RI = 12.0% ± 6.8% and 66.0% ± 12.3%,respectively).h-GCC colonies also showed typical adenocarcinomatous glandular structures around the h-hepatocyte-colonies.These mice survived for the full 56 day-study and did not exhibit any metastasis of h-GCCs in the extrahepatic regions during the observational period.The mice with an h-hepatocyte-repopulated liver possessed metastasized h-GCCs and therefore could be a useful humanized liver animal model for studying liver cancer metastasis in vivo.CONCLUSION:A novel animal model of human liver cancer metastasis was established using the uPA/SCID mouse line.This model could be useful for in vivo testing of anti-cancer drugs and for studying the mechanisms of human liver cancer metastasis.

OPN基因沉默抑制MKN28细胞生长、侵袭及凋亡

目的检测骨桥蛋白(osteopontin,OPN)对人胃癌细胞MKN28生长、侵袭及凋亡的影响。方法利用脂质体转染的方法 ,进行胃癌转移相关基因骨桥蛋白(OPN)的siRNA,并通过荧光定量PCR以及免疫荧光的方法 ,进行siRNA效果鉴定。根据siRNA的抑制效果,将细胞分为正常对照组、非特异干扰组、特异干扰24h组、特异干扰36h组和特异干扰48h组,比较对照组与不同siRNA作用时间组的细胞增殖、细胞凋亡、迁移黏附能力的改变。结果 siRNA作用48h组,OPN的表达最低;相应地,其细胞活力降低,增殖抑制率达30.20%,迁移黏附能力降低,基质膜上的细胞数目明显减少[对照组与OPN干扰组比较:(42.0±9.4)vs(21.8±6.9),P<0.01],细胞对基底膜基质和纤维连接蛋白的黏附百分比明显降低[对照组与OPN干扰组比较:基底膜基质,(41.5±8.4)%vs(20.5±4.5)%;纤维连接蛋白,(25.3±4.5)% vs(14.6±2.5)%],凋亡增加。结论 OPN基因沉默能有效抑制人胃癌细胞MKN28的生长、侵袭,并促进凋亡的发生。