Objective To establish a new model of orthotopic-transplatation tumor of human malignant choroidal melanoma. Methods pEGFP-N1, the eukaryotic expressive plasmid of green fluorescent protein (GFP), was transfered into human malignant choroidal melanoma cell line (OCM-1) by liposome lipofectanine, then the cell clones with stable GFP expression were screened out by means of neomycin, fluorescence microscope, and flow cytometer. Two μl cell suspension of OCM-1 cells with GFP expression with the density of 4.5×107-5.5×107 cells/ml was injected into the subretinal space of right eyes of 40 nude mice (40 eyes) under binocular operating microscope with left eyes as the control ones. The growth of the transplanted malignant choroidal melanoma was observed in vivo using the fluorescence stereomicroscope. The mice were killed at different time after the operation to observe the metastasis of the tumor to optic nerve, brain and other organs including lung, liver, kidney and spleen. Moreover, pathological detection and immunohistochemical staining of GFP was carried out. Results At the postoperative 10th-12th days, the growths of the transplanted malignant choroidal melanoma with dilated and distorted blood vessels and neovascularization were observed; at the postope rative 20th-22nd days, the melanoma occupied the whole cavity of vitreous body; and at the postoperative 24th-26th days, the transplanted tumor grew out of the eye. Metastases of the carcinoma to olfactory bulb, kidney, lung and liver were seen at the failure phase soon after the extra-ocular phase. The histopathological characteristics of the transplanted tumor were similar to those of human, and the results of immunohistochemical staining showed positive expression of GFP in the tumor cells. Conclusion The orthotopic model of malignant choroidal melanoma set up via injection of human malignant choroidal melanoma cells labeled by GFP into the subretinal space of nude mice may provide a new approach to investigate the natural courses of growth and metastasis of malignant choroidal melanoma. (Chin J Ocul Fundus Dis，2004，20：245-248)
ObjectiveTo explore the relation between vascular endothelial growth factor （VEGF） and the formation of tumor thrombosis in the main trunks of portal vein (PVTT). MethodsTumor specimens were collected from 36 patients (16 patients with PVTT, the other patients without PVTT and metastasis) undergoing resection of hepatocellular carcinoma (HCC) and portal thrombectemy, PVTT specimens of 16 patients named group A1, the same patients’ with HCC named group A2, tumor specimens of the other patients named group B. In situ hybridization and immunohistochemistry were used to investigate VEGF mRNA, protein and microvessel density (MVD) on surgical specimens. The intensity was evaluated using a computer image analyzercell analysis system.ResultsVEGF mRNA expression was detected in the tumor’ cell of the specimens. The expression rates of VEGF mRNA in the group B, A2, A1 were 30%, 100%, 100% respectively, and the expression rates of VEGF mRNA in group A2 and A1 were higher than that in group B （P＜0.01）. The intensity of VEGF mRNA in group A2 （0.078 5±0.019 6） were lower than in group A1 （0.194 4±0.059 0） （P＜0.01）. VEGF protein expression was often detected in the tumor cell, vascular endothelial cell and fibroblast cells. Invasion was detected in small vein in group A2, more tumor cell colony detected in group A1. The expression rates of VEGF protein in group B, A2, A1 were same as VEGF mRNA; the intensity of VEGF protein in A1 （0.165 6± 0.034 5） was higher than in group A2 （0.108 1±0.024 3） （P＜0.01）. MVD in group B, A2, A1 was 31.9±14.4, 63.3±15.1, 116±27.6/view of 200 microscopefield, MVD in group A1 was higher than group A2 （P＜0.01）, higher in group A2 than in group B. There was a statistically significant correlation between the intensity of VEGF expression and MVD in group B,A2 and A1. ConclusionVEGF could play an important role in the invasion, metastasis of HCC and the formation of PVTT. Angiogenesis in tumor is correlated well with the progression of HCC.
Objective To investigate the invasion ability of Panc-1 cells in vivo and in vitro af ter being t ransfected with tissue factor pathway inhibitor 2 gene ( TFPI-2) . Methods The expression vector pEGFP-C1-TFPI-2 was transfected into human pancreatic cancer line Panc-1 cells by using liposome. TFPI-2 mRNA and protein of transfected and nontransfected cells were detected by reverse t ranscription-polymerase chain reaction (RT-PCR) and Western blot respectively. The tumor cells invasive behavior of t ransfected ( Panc-1-TFPI-2) and nontransfected ( Panc-1-V and Panc-1-P) cells were assessed in vitro through Boyden Chamber method. The transfected and nontransfected cells were implanted into nude mice to observe it s growth and metastasis in vivo. Results Expressions of mRNA and protein of TFPI-2 were confirmed in transfected cells. Af ter TFPI-2 t ransfection , the number of Panc-1-TFPI-2 , Panc-1-V and Panc-1-P cells passing through membrane of Boyden Chamber were 24. 4 ±3. 5 ,61. 3 ±4. 1 and 60. 2 ±3. 9 , respectively. The number of TFPI-2-expressing cells to t raverse a Matrigel-coated membrane was obviously decreased compared with that of non-expressing cells , the invasion ability was lower than that before transfection in vitro. The subcutaneous tumor volume of the Panc-1-TFPI-2 group was (438. 0 ±69. 8) mm3 , the Panc-1-V group was (852. 0 ±102. 9) mm3 and the Panc-1-P group was (831. 0 ±78. 1) mm3 , P lt; 0. 05. The metastasis to liver and lung and muscular invasion occurred in the Panc-1-V group and the Panc-1-P group. There were no muscular invasion and metastatic lesions in the Panc-1-TFPI-2 group. Conclusion TFPI-2 gene expression may obviously inhibit the invasion ability of pancreatic cancer cells in vitro and in vivo , which provides an experimental basis for the treatment of human pancreatic cancer by gene therapy.
ObjectiveTo evaluate the prognostic significance of metastatic lymph nodes ratio (MLNR) in patients with node-positive breast cancer. MethodsThe clinical data of 94 patients with nodepositive breast cancer underwent modified radical mastectomy were retrospectively analyzed. The survival rate and prognosis factors of patients with complete follow-up data were assessed by log-rank test and multivariate regression analysis. Results The survival time of 94 patients ranged from 12-75 months, with median 64 months. The 5-year overall survival rate was 72.34% (68/94). The total MLNR was 0.31 (486/1 553). Univariate analysis demonstrated that the survival was influenced significantly by tumor size, number of lymph node metastasis, MLNR, ER status, and radiotherapy or not (Plt;0.05), but not by patient’s age, menopause or not, PR status, endocrine therapy or not, and histological type (Pgt;0.05). Multivariate analysis showed that MLNR (OR=2.565, 95%CI=1.043-6.309， P=0.040) and tumor size (OR=2.220, 95%CI=1.045-4.716， P=0.038) were independent prognostic factors for the patients with node-positive breast cancer. Conclusion MLNR is a major independent prognostic factor for the patients with node-positive breast cancer, which is more accurate than the number of metastatic lymph nodes in predicting the survival of patients with node-positive breast cancer.
Objective To investigate the influence of CO2-insufflation pressure on invasion potential of the colon cancer cells. Methods With an in vitro artificial pneumoperitoneum model, SW1116 human colon cancer cells were exposed to CO2-insufflation of 5 different pressure groups: 6, 9, 12, 15 mm Hg and control group, respectively for 1 h. The invasion capacities of SW1116 cells exposed to CO2-insufflation of 5 different pressure groups were detected by cell adhesion/invasion assay in vitro. Results Immediately following exposure to 15 mm Hg CO2 insufflation, the invasion of SW1116 cells decreased significantly compared to the cells before exposure. At the 0 h time point, the cells exposed to 15 mm Hg were significantly less invasive than those exposed to the other insufflation pressure (P＜0.05), and the cells exposed to 6 mm Hg were more invasive than cells exposed to the other insufflation pressure (P＜0.05). And 72 h after exposed to CO2-insufflation, the differences between the pressure groups were not significant. Conclusion CO2-insufflation induced a temporary change in the invasion capacity of cancer cells in vitro, higher pressure of CO2-insufflation inhibits the invasion potential.
Objective To approach histologically the degree of choroidal invasions of retinoblastoma and their relations with metastasis and prognosis. Methods The pathological sections of 297 enucleated eyeballs with retinoblastom from June 1985 to 1995 were reviewed under light microscope.For the sake of assessing the risk factors of the prognosis of the disease,the graduation of the choroidal invasion was in vestigated in particular,and the follow-up of this genesis were performed as well. Results According to the degree of affection of the intraocular tissues from the tumor cells especially the choroid as the crucial structure,the choroidal invasions were classified into the following 5 categories:0，prereactive phase of retinal pigment epithelium(144/297,48.48%).Ⅰ，reactive phase of retinal pigment epithelium(81/297，27.27%).Ⅱ, early phase of choroidal invasion( 29/297,9.76%).Ⅲ, middle phase of choroidal invasion(17/297,5.72%).Ⅳ， advanced phase of choroidal in vasion(26/297,8.75%).24.24% of cases were found in phases Ⅱ-Ⅳ (so called choroidal invasion).The mortality in patients with phases 0～Ⅲ was found to be 0.4% in the average follow-up period of 51.8 months,and inpatients with phase Ⅳ was found to be 12.0%. Conclusion The retinoblastoma patient with the advanced phase of choroidal invasion (choroida l invasion with great extent) had relative high metastatic rate and poor prognos is. (Chin J Ocul Fundus Dis,1999,15:88-90)
ObjectiveTo systematically review the efficacy and safety of 89SrCl2 for bone metastases.MethodsDatabases including PubMed, EMbase, The Cochrane Library (Issue 12, 2016), MEDLINE, CBM, VIP, CNKI and WanFang Data were electronically searched from inception to December 2016 to collect randomized controlled trials (RCT) about 89SrCl2 in the treatment of bone metastases. Two reviewers independently screened literature, extracted data, and assessed the risk of bias of included studies. Then meta-analysis was performed by using RevMan 5.3 software.ResultsA total of 19 RCTs involving 1 899 patients were included. The results of meta-analysis showed that: compared to the group without 89SrCl2, group of 89SrCl2 combined with other treatments had benefits for complete relief of bone pain (RR=1.53, 95%CI 1.16 to 2.01, P=0.002), total relief of bone pain (RR=1.33, 95%CI 1.22 to 1.45, P<0.001), bone metastases complete response rate (RR=1.61, 95%CI 1.20 to 2.16, P=0.002), bone metastases total response rate (RR=1.61, 95%CI 1.26 to 2.08, P<0.001), improvement of quality of life (RR=1.44, 95%CI 1.06 to 1.97, P=0.02). The proportion of pain flare (RR=10.35, 95%CI 2.01 to 53.37, P=0.005), leucopenia (RR=2.49, 95%CI 1.84 to 3.37, P<0.001) and thrombocytopenia (RR=2.64, 95%CI 1.76 to 3.97, P<0.001) in the 89SrCl2 group were higher than that in the control group.Conclusion89SrCl2 combined with the other treatments is better for bone metastases than without 89SrCl2. Due to the limited quantity and quality of included studies, the above results should be verify by more high quality studies.
Objective To evaluate the therapeutic effects of percutaneous microwave ablation (MWA) for metastatic liver cancer. Methods Ultrasound-guided percutaneous MWA technique was used to treat 27 cases of hepatic metastases with 69 nodules 〔0.9-13.2 (3.0±2.0) cm in diameter〕. Local therapeutic effect, local and distant recurrence rate and survival rate were evaluated respectively. Results The complete ablation (CA) rate which was used to evaluate the local therapeutic effects was 92.8% (64/69), with 100% (34/34), 92.3%(24/26) and 66.7% (6/9) in a diameter of nodules lt;3.0 cm, 3.0-5.0 cm and ≥5.0 cm respectively. The CA rate was lower in the group of a diameter of nodules ≥5.0 cm as compared with other two groups (Plt;0.05). The local recurrence rate was 9.4% (6/64), with 2.9% (1/34), 16.7% (4/24) and 16.7% (1/6) in a diameter of nodules lt;3.0 cm, 3.0-5.0 cm and ≥5.0 cm respectively. The intrahepatic distant recurrence rate was 44.4% (12/27). The follow-up time after MWA was 3－34 (17.0±8.7) months. During the follow-up period, 6 months, 1 and 2-year cumulative survival rate was 88.9%, 63.0% and 34.4% respectively, with a mean survival time of 17.8 months, and with a median survival time of 19.0 months.Conclusion Percutaneous MWA treatment offers satisfactory local tumoricidal efficacy to metastatic liver cancer and the patients with recurrence and new metastases can be therapy repeatedly to improve long-term survival.
ObjectiveTo observe the clinical features of uveal metastases from lung carcinoma.MethodsA retrospective case study. From 1983 to 2014, 14 patients with uveal metastases of lung cancer confirmed by ocular examination in Peking Union Medical College Hospital were included in the study. Among them, 7 were male, 7 were female; 11 were monocular and 3 were binocular. The mean age was 54.5±9.6 years. Pathologic examination showed primary bronchial lung cancer, including 13 patients of non-small cell lung cancer (10, 2 and 1 patients of lung adenocarcinoma, squamous cell carcinoma and adenosquamous cell carcinoma, respectively) and 1 patient of small cell lung cancer. Four patients (28.6%) were diagnosed with lung cancer before ophthalmology consultation, and 10 patients (71.4%) were first diagnosed with ophthalmology due to ocular symptoms. The duration from ocular symptoms to lung cancer diagnosis was 1 week to 6 months. The course from diagnosis of lung cancer to ophthalmological consultation was ranged from 10 to 60 months, and the average course was 29.5±19.0 months. There were 7, 4 and 3 patients with impaired vision, occlusion of visual objects and deformation of visual objects, respectively. All patients underwent visual acuity, slit lamp microscope, B-mode ultrasound and UBM examinations. FFA was performed in 8 eyes, and 2 eyes were examined for ICGA. Orbital MRI was performed in 5 patients. Vitreoretinal surgery was performed on 1 eye. The clinical characteristics of the patients were analyzed and observed.ResultsIn 17 eyes, there were 2 eyes with visual acuity of light perception, 3 eyes of hand movement to counting finger before the eyes, 5 eyes of 0.1-0.3, 4 eyes of 0.4-0.6, 3 eyes of greater than 0.8. Metastatic cancer was located in iris in 1 eye, it presents as a red mass with irregular shape on the surface, which is full of small nourishing blood vessels. Metastatic cancer were located in choroid in 16 eyes, they presented yellowish-white or grayish-yellow lumps under the posterior pole or equatorial retina, including 14 eyes with a single lesion and 2 eyes with 2 lesions, with retinal detachment in 8 eyes and increased intraocular pressure in 5 eyes. B-mode ultrasonography showed posterior polar flat or surface irregular wavy intraocular space occupying lesions with localized or extensive retinal detachment. FFA and ICGA showed the focal, apical and patchy fluorescence of the tumor. MRI showed that T1WI medium and high signal consistent with the vitreous body, while T2WI showed low signal.ConclusionsUveal metastatic may be the first manifestation of lung cancer, and visual impairment, part of solid mass lesions with fundus flattening may be accompanied by secondary glaucoma and retinal detachment as the main clinical manifestations. Most of the metastatic sites are located in choroid, which is more common in single eye and single lesion. Adenocarcinoma is the most common type of uveal metastasis in non-small cell lung cancer.
Objective To investigate the influence of different pressures and duration of CO2 pneumoperitoneum on the adhesive and invasive ability of gastric cancer cells based on the expressions of adhesive and invasive molecules. Methods With an artificial CO2 pneumoperitoneum model in vitro, human gastric cancer cell lines including MKN-45, SGC-7901, and MKN-28 were exposed to CO2 in different environments: 0 mm Hg （1 mm Hg=0.133 kPa）, 9 mm Hg （2 h, 4 h）, and 15 mm Hg （2 h, 4 h）. The expressions of mRNA of E-cadherin, intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase-2 (MMP-2), and vascular endothelial growth factor-A (VEGF-A) in the different environments were measured by RT-PCR. The expressions of protein of E-cadherin and ICAM-1 in the environments of 0 mm Hg and 15 mm Hg （4 h） were measured by FCM. Results With the increase of duration or pressure, RT-PCR showed that there was a downward trend in the expression of E-cadherin mRNA as well as there were upward trends in the expressions of ICAM-1, MMP-2, and VEGF-A mRNA; FCM showed that there was a downward trend in the expression of E-cadherin protein while the expression of ICAM-1 protein showed the opposite change. But there were no obvious differences under different environment (P＞0.05). Conclusions Under low pressure (≤15 mm Hg) and short time (≤4 h) of CO2 pneumoperitoneum, the adhesive and invasive ability of gastric cancer cells could not be affected, which means that under this environment, CO2 pneumoperitoneum will not increase the possibility of neoplasm metastasis.