Objective To investigate the effects of adenovirus-mediated melanoma differentiation-associated gene-7 (mda-7)/IL-24 and/or adriamycin (ADM) on transplanted human hepatoma in nude mice and to explore a new way for hepatoma gene therapy combined with chemotherapy. Methods The recombinant adenovirus vector carrying Ad.mda-7 was constructed; Ad.mda-7 and/or ADM were injected into the tumor-bearing mice. Their effects on the growth of the tumor and the survival time of the mice were observed. The expressions of VEGF and TGF-β1 were detected by an immunohistochemistry method. Results Ad.mda-7 was constructed and expressed in vivo successfully. Compared with other three groups 〔control group (43.4±1.67) d, ADM group (64.2±4.14) d, Ad.mda-7 group (61.4±1.67) d〕, the mice treated with Ad.mda-7 combined with ADM had longer average survival time 〔(83.8±4.82) d, P＜0.01〕; the average size of tumor treated with Ad.mda-7 combined with ADM diminished significantly compared with that treated with ADM or Ad.mda-7 separately (P＜0.01). VEGF and TGF-β1 expressions of Ad.mda-7 group were (56.2±7.7)%, (35.2±4.5)%, respectively, and were lower than those in ADM group (VEGF: P＜0.05; TGF-β1: P＜0.01). VEGF expression of Ad.mda-7+ADM group was (37.3±5.0)%, and was significantly lower than that in other three groups (P＜0.01). TGF-β1 expression of Ad.mda-7+ADM group was (31.2±3.1)% and significantly lower than that in control group and ADM group (P＜0.01), however, there was no significant difference compared with Ad.mda-7 group (Pgt;0.05). Conclusion Ad.mda-7 combined with ADM has b antitumor potency and synergistic effects and suppresses the growth of human HCC xenograft in nude mice, possibly by inducing the apoptosis of hepatoma cell lines and suppressing tumor angiogenesis.
Objective To investigate the role of anti apoptosis gene bcl-2 in the survival of cultured uveal melanoma UM cells. Methods Antisense oligodeoxynucleotides (AS-ODN) bcl-2 were delivered with cationic lipid to primary cultured UM cells. The inhibitory effect of AS-ODN bcl-2 on proliferation of UM cells was examined by 3,-4,5 Dimethyliazol-2,5 diphenyl tetrazolium bromide (MTT) method. Using DNA ladder to determine if the UM cells had been apoptotic. Bcl-2 expression was detected by RT-PCR and Westernblot technics. Results The effect of AS-ODN bcl-2 in inhibiting the proliferation of cultured UM cells had opposite relation to dosage. It down regulated the mRNA and protein level of bcl-2 gene, and the sense ODN didn′t have this effect. Conclusion AS-ODN bcl-2 can down regulate bcl-2 expression, inhibits UM cells proliferation and induces apoptosis. (Chin J Ocul Fundus Dis, 2002, 18: 38-41)
Intraocular tumors is a serious blinding eye disease, which has a serious impact on patients' vision and even life. At present, the main treatments include surgical treatment, radiation therapy, chemotherapy, laser therapy and combination therapy. In recent years, with the wide application of anti-vascular endothelial growth factor (VEGF) in the treatment of ocular diseases, many studies have confirmed that anti-VEGF drugs play an important auxiliary role in the treatment of intraocular tumors and its complications. In terms of the therapeutic effect, intravitreal anti-VEGF combined with other methods have a good prognosis in the treatment of choroidal metastatic carcinoma and retinoblastoma, while the therapeutic effect of uveal melanoma is still controversial. In the treatment of intraocular tumor complications, intravitreal anti-VEGF also has a good effect on the secondary lesions of choroidal osteoma and radiation retinopathy. As for drug safety, intravitreal anti-VEGF can significantly reduce the toxic and side effects of systemic chemotherapeutic therapy. However, the dosage and medication regimen of anti-VEGF drugs in the treatment of intraocular tumors and their complications have not been unified in current studies, and further basic and clinical trials are still needed to explore in the future.
Objective To investigate the effects of matrine on proliferation, apoptosis and radiotherapy sensitivity of uveal melanoma cells. MethodsAn animal experiment study. In vitro experiment: MuM2B cells of human choroidal melanoma were randomly divided into control group and matrine 0.25, 0.50, 1.00, 2.00 g/L groups. The cell morphology was observed by transmission electron microscope. Cell proliferation was detected by thiazole blue colorimetry. The mRNA and relative expression levels of CyclinD D (CyclinD), B lymphoblastoma-2 (Bcl-2) and Bcl2-associated X protein (Bax) were detected by real-time polymerase chain reaction and Western blot. In vivo experiment: BALB/C mice were injected with MuM2B cell suspension subcutaneously on the back of forelimb to prepare transplanted tumor model. After successful modeling, they were randomly divided into blank group and matrine treatment group with different concentrations. Mice in blank group were injected with phosphate buffer subcutaneously. Mice in different matrine treatment groups were injected with 15, 25, 50, 100 mg/kg matrine subcutaneously, respectively, for 7 consecutive days. The tumor was weighed and its volume was measured after the last administration. Single factor analysis of variance was used to compare different groups. The t test was used for pairwise comparison between groups. ResultsIn the control group, the cell structure was normal, the distribution was uniform, and no or rare nuclear pyknosis was seen. With the increase of matrine dosage, the nuclear pyretosis increased gradually and cell morphology changed obviously. Compared with the control group, the cell survival rate in 0.50, 1.00 and 2.00 g/L groups gradually decreased with matrine concentration increasing and treatment time prolongating, the relative expression levels of CyclinD and Bcl-2 mRNA and protein gradually decreased, and the relative expression levels of Bax mRNA and protein gradually increased. Under the same radiation dose X-ray irradiation, the cell survival rate of 0.50, 1.00 and 2.00 g/L groups gradually decreased, and the differences were statistically significant (P＜0.05). Compared with blank group, the tumor weight and volume of mice in different doses of matrine group were significantly decreased, and the differences were statistically significant (P＜0.05). ConclusionMatrine can down-regulate the expression of CyclinD and Bcl-2, up-regulate the expression of Bax, promote the apoptosis of MuM2B human melanoma cells, inhibit cell proliferation, and enhance cell radiosensitivity.
Objective To establish an allogenic intraocular melanoma model and observe its pathological features.Methods Thirty-six kunming mice were devided randomly into 3 groups with 12 ones in each, and allogeneic melanoma cells B16F10(C57BL16) were inoculated into the anterior chamber (AC), vitreous cavity (VC) of right eyes and under the skin (subcutaneous, SC) of the back of right feet of each grup respectively. The incidence of tumor occurance, time of breaking through the eyeball and other general pathologic features of the tumor were observed by slip-lamp biomicroscopy and operating microscopy for continuous 32 days, and the results were statistically analyzed. Pathological examination was given for tumors at last.Results The incidence of tumor occurance in both AC (12/1 2 eyes) and VC group (11/11 eyes) was higher than that in SC group (2/12 feet)（χ2=17.143, P＝0 .000；χ2=16.218, P=0.000）. The time of eyeball diabrosis was 11-13 days in AC group and 13-32 days in VC group, and there was significant difference between these two groups (Log Rank=18.22, P=0.000). The intraocular melanomas could grow progressively, but reduced and fell off when they broke through eyeball and grew in or bit for a period. The average diameter of the tumor after 32 days after inoculation was (2.27±1.97) mm in AC group，(3.82±1.85) mm in VC group and (0.94±2.27) mm in SC group. There was significant difference between VC and SC group (t=3.322，P=0.003). In pathohistological examination, tumor tissue necrosis could be observed at the center of the subcutaneous melanomas but not in intraocular melanomas.Conclusions Allogeneic intraocular melanoma model is successfully established which is convenient, repeatable, and helpful to studying the mechanism of genesis and development of this tumor. (Chin J Ocul Fundus Dis，2003，19：333-404)
PURPOSE:To evaluate the value of the apoptosis-suppressing oncogene bcl-2 protein expression in the development and progression of uveal and conjunctival melanomas.
METHODS:Using flow cytometry and immunofluorescence methods to detect the bcl-2 protein expression in 40 cases of uveal malignant melanomas (UMM), 5 cases of conjunctival nevi (CN) and 7 cases of conjunctival malignant melanomas (CMM).
RESULTS :The expression content of bcl-2 protein in CMM was significantly higher than that in CN (P＜0.05);the bcl-2 protein positive expression percentages in CMM and UMM were 85.71% and 72.50% respectively. The expression content of bcl-2 protein in UMM was not related to pathological
classfication, scleral invasion,ciliary body involvement,and tumor dimensions (P＞0.05).
CONCLUSIONS: The over-expression of bcl-2 protein and apoptosis suppressing might be related to the pathogenesis of CMM and UMM;bcl-2 protein expression might be helpful in discriminating CN from CMM, but unavailable in evaluating the patholgical malignancy of UMM.
（Chin J Ocul Fundus Dis,1997,13: 73-74 ）
Objective To observe the macular morphological changes of choroidal melanoma with coherence tomography (OCT) after plaque radiotherapy (PRT). Methods A total of 48 patients (48 eyes) with choroidal melanoma who underwent125I PRT were enrolled in this study. All the patients were examined documenting OCT to get the image of macula. The macula of all the patients was not involved. The median visual acuity was 0.4plusmn;0.2, which ranged from 0.02 to 1.0. There were 18 eyes (37.5%) with retinal detachment, 12 eyes (25.0%) with retinal pigment epithelium (RPE) changes, seven eyes (14.6%) with macular edema, epimacular membrane, detachment combined with edema, exudation and RPE changes, 11 eyes (22.9%) with normal macular structure. The median follow-up time was (10.4plusmn;5.9) months, which ranged from one to 24 months. The tumor control situation and visual acuity were observed in follow-up period. The same equipment and methods of OCT were used to return visit in follow-up period. The macular morphological changes at the final visit and its relationship with PRT and visual acuity were contrastively analyzed. Results All the patients had good control of tumor. The vision acuity improved in two eyes (4.2%), unchanged in 10 eyes (20.8%), and decreased in 36 eyes (75.0%). The differences of the visual acuity was statistically significant between before and after treatment (Z=-3.778,P＜0.05). There were 13 eyes (27.1%) with retinal detachment; nine eyes (18.8%) with RPE changes; 17 eyes (35.4%) with macular edema, detachment combined with edema, exudation and RPE changes; six eyes (12.5%) with proliferation, atrophy, detachment combined with edema, exudation and epimacular membrane;three eyes (6.3%) with normal macular structure. There were 15 patients (31.3%) with two or more abnormal macular morphology after PRT. Conclusions Retinal detachment, RPE changes, macular edema and exudation are common abnormal macular morphology after PRT. The incidence rate of abnormal macular morphology is increased. There are 31.3% patients with two or more abnormal macular morphology.
Objective To observe the different clinical response patterns of uveal melanoma (UM) patients after external scleral plaque radiotherapy (PRT), and to investigate the risk factors of secondary enucleation after treatment failure. MethodsA single-centre retrospective study. Demographic baseline characteristics and clinical data were collected from 465 UM patients treated with 125I external scleral PRT at Beijing Tongren Hospital from March 2011 to September 2017. Among them, 217 were male and 248 were female, tumor all occurred monocularly. The mean age of subjects was 46.7±12.1 years. Reasons for secondary enucleation included local tumor treatment failure, glaucoma, scleral necrosis and patient request. Tumor grading was based on the grading standards established by the American Joint Committee on Cancer (AJCC). The pattern of tumor response after PRT was classified as degenerated type, growth type, stable type or other types according to literature criteria. The median follow-up time after PRT was 59 months to observe tumor changes. Complete follow-up records of 3 or more color doppler ultrasound imaging (CDI) was available in 245 cases. A t-test was performed to compare the patient's age, intraocular pressure, best corrected visual acuity, tumor thickness and maximum basal diameter before treatment; a chi-square test was performed to compare the patient's gender, AJCC T classification of the tumor, whether the ciliary body was involved, presence of subretinal fluid, optic disc invasion and vitreous hemorrhage, tumor shape and location. Kaplan-Meier survival analysis was used to estimate the cumulative probability of secondary enucleation after extra-scleral PRT. Univariate and multivariate Cox proportional hazards regression analyses were used to evaluate the relationship between tumor characteristics and secondary enucleation after extra-scleral PRT. ResultsAmong 465 patients, eecondary enucleation was performed on 78 (16.8%, 78/465) patients during the follow-up period. The 1, 3 and 5 year secondary enucleation rates were 5.4%, 9.3% and 17.1%, respectively. Eye preservation was successful in 387 cases (83.2%, 387/465). Patients treated by secondary enucleation had a larger maximum basal diameter of tumor, a higher proportion of irregular and diffuse morphology, a cumulative macular involved and a higher AJCC T classification, the difference was statistically significant (P＜0.05). There were 115, 76, 27, and 27 cases of degenerated type, stable type, growth type, and other type, respectively. The tumor thickness of the growth type and other types was significantly smaller than that of the degenerated type and the stable type, and the difference was statistically significant (P＜0.05). Univariate Cox analysis showed that the maximum basal diameter of the tumor (HR=1.19), tumor thickness (HR=1.08), AJCC T classification (HR=1.90), growth type response pattern (relative to degenerated type response pattern) (HR=4.20) was associated with failure of eye preservation (P＜0.05). In the multivariate Cox analysis, the largest tumor basal diameter (HR=1.24) and the growth type response pattern (relative to the degenerated type response pattern) (HR=4.59) were still associated with failure of eye preservation (P＜0.05). ConclusionsThe tumor thickness of UM patients with growing and other response patterns after PRT is smaller before treatment; the maximum basal diameter of the tumor and the growing response pattern are independent risk factors for secondary enucleation.
【Abstract】ObjectiveTo detect the expression of melanoma antigen-3 gene (MAGE-3) in rectal cancer and explore its relationship with the clinical pathology of rectal cancer and clinical significance in immune therapy. MethodsThe expressions of MAGE-3 in tumor tissue, para-tumor tissue of （5±1） cm from border of tumor tissue, tissue of resection border and polypus were detected by RTPCR. ResultsMAGE-3 expression was positive in 14 of 33(42.42%) samples of rectal cancer, 6 of 33 (18.18%) of paratumor tissue, and 5 of 33 (15.15%) of resection border tissue respectively. No MAGE-3 was detected in 3 polypi. The positive rate of MAGE-3 in tumor tissue was significantly higher than that of the paratumor and resection border tissue (P＜0.05). The expression of MAGE-3 was not related to age, gender, histological type, metastasis to lymph nodes, and Dukes stage (Pgt;0.05). Conclusion MAGE-3 protein may be used as a target of immunotherapy for rectal cancer and an index for follow-up and screening of rectal cancer.
Objetive
To observe the incidence of extraocular extension of malignant uveal melanoma, the relationship between the incidence and histopathologic type, the characters of clinical manifestation and iconography, and the factors influencing prognosis.
Methods
Nine cases of extraocular extension of malignant uveal melanoma within last 20 years were analyzed retrospectively. All cases were diagnosed by histopathological examination and most of them had undergone ultrasound and CT examinations.
Results
The incidence of extraocular extension of malignant uveal melanoma was 15%. Tumors of 6 patients were flat and 2 nodular in shape, 4 epithelial cell and 1 spindle cell in type. The follow-up results of 5 patients in this series indicated that the prognosis was related to the size of the tumor, the degree of extraocular extension, and histopathologic type.
Conclusion
Extraocular extension of malignant uveal melanoma might happen in early case and the occurrence is usually related to the histopathologic type of the tumor. Iconographic examination is very helpful for diagnosis in extraocular extension of this tfumor.
(Chin J Ocul Fundus Dis,1999,15:30-32)
