Retinoblastoma (RB) is the most common intraocular malignant tumor in children. With advanced clinical technologies there are more and more methods available to treat retinoblastoma, and make it is possible to delivery individualized protocol combined traditional treatments with modern regimen to patients now. In order to improve the survival rate and the life quality of RB patients in China, it is very important to make a suitable system of standardized therapy based on results from developed countries and health policies of our own country.
ObjectiveTo evaluate the efficacy of intra-arterial chemotherapy (IAC) for advanced retinoblastoma (RB) after failure of intravenous chemotherapy (IVC). MethodsFifteen eyes of 13 patients with advanced RB were treated with IAC (1-5 cycles) after failure of IVC (2-8 cycles). The patients included 10 boys and 3 girls, with the mean age of (15.67±8.16) months. Six patients had bilateral RB and 7 patients had unilateral RB. There were 14 eyes (93.33%) in stage D, 1 eye (6.67%) in stage E according to the International Classification of intraocular retinoblastoma. The main reasons for failure of IVC were recurrent primary tumor in 3 eyes (20.00%), subretinal seeds recurrence in 9 eyes (60.00%), viable vitreous seeds in 2 eyes (13.33%) and poor response of primary tumor in 1 eye (6.67%). The mean interval between IVC completion and IAC start was 3 months. The mean follow-up was 19 months (ranged from 3 to 52 months). ResultsAfter IVC and secondary IAC, the retinoblastoma and seeds were regressed in 12 eyes (80.00%). Three eyes required enucleation for severe vitreous seeds, subretinal seeds recurrence and primary tumor recurrence. There was no evidence of metastasis in any case. ConclusionIAC can achieve high global salvage rate (80.00%) for patients with advanced retinoblastoma after failure of IVC.
Objective To investigate the effect of doxycycline on the proliferation and vasculogenic mimicry in retinoblastoma (RB) cell line in vitro. Methods RB cell line were tested for their ability to form perfusable tubular networks in 3D culture with doxycycline in the concentrations ranging from 5 to 20 mg/L, and CoCl2 was used as chemical hypoxia-inducing reagent to mimic tumor hypoxic microenvironment. The effect of doxycycline on proliferation were detected by MTT assay in vitro, and the effect on tube formation of RB cells were detected by tube-like structure formation assay and PAS staining. The mRNA levels of MMP2 and MMP9 at different hypoxic culture and different doxycycline concentrations were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Results The micrograph showed that RB cells linked each other to form cavity and network tructure in 3D culture. the number of tubules in doxycycline group were significantly lower than which in the control group in the concentrations ranging from 5 to 20 mg/L (Plt;0.001).OD of doxycycline group was significantly lower than which in the control group (t=15.320，Plt;0.01) , The proliferation of RB cells had a negative correlation with the concentration of doxycycline (r =-0924, Plt;001). The levels of MMP2 and MMP9 mRNA of RB cells under hypoxia were significantly higher than which in the control group (t=16.469，Plt;0.01). As the concentration of doxycycline increased, the expression of MMP-2 and MMP-9 decreased. The result of double staining also showed that VM, formed by CD34negative and PASpositive tumor cells, existed in 12 simples of retinoblastoma. Conclusion RB cells have the capacity of selfmetamorphosing and vasculorizing in 3D culture. Doxycycline can inhibit their proliferation and vasculogenic mimicry formation in vitro by downregulating the expression of MMP-2 and MMP-9 .
Thanks to the treatment of retinoblastoma (RB) having improved significantly in recent years, there is an increasing trend to use conservative treatment modalities that aim to preserve the globe as well as vision with minimum mortality. RB therapy is a long-term systemic treatment in clinical practice. Although there are many treatment options for RB therapy, such as cryotherapy, photocoagulation, systemic chemotherapy, enucleation and ophthalmic chemotherapy, it is recommended to consider in accordance with the following key points in gaining a reasonable treatment strategies: to make sure that RB is an intraocular period; to determine whether the intraocular RB to be treated with eye preservation or enucleation; what is the case of eye preservation therapy combined with chemotherapy and how to arrange the follow-up of RB patients. It's more complicated to choice the therapeutic measures for RB in clinical practice. So, the patient's condition, economic capability and medical condition should be evaluated comprehensively. The principle of RB treatment should be followed, which is protecting eyeball and visual function without life damage.
Objective A short-term observation study of ocular surgery with or without systemic chemotherapy to treat retinoblastoma (RB). Method The clinical data of 66 RB cases treated at Beijing Tongren Hospital from August 2006 to September 2007 were retrospective reviewed.All patients received ultrasound (B scan/CDI), radiology examination (CT scan/MRI) and RetCam retina examination, and classified according to the International Intraocular Retinoblastoma Classification (IIRC) method. Focal therapies (include laser and cryotherapy) were applied to Group A, systemic chemotherapy (CCTV protocol, Cyclosporine, Carboplatin, Teneposide and Vincrinstine) plus ocular surgery (include focal therapy, enucleation and orbital exenteration) to Group B, C, D and E. Patients were followedup for more than 6 months. Result Of 66 cases (36 male/30 female), 16/66 cases were bilateral RB and 50/66 were unilateral. Follow up period was from 6 to 13 months (mean time 8.6 month). According to IIRC, there were 5 eyes in Group A, 6 eyes in Group B, 5 eyes in Group C, 15 eyes in Group D and 51 eyes in Group E( include extraocular disease). 22/82 eyes were conserved at the end of follow up, including 5/5(100%) of group A, 6/6(100%) of group B, 5/5(100%) of Group C, 4/15(26.7%) of Group D and 2/51(3.9%) of Group E. 5/66 (7.6%) patients died during this period, they all were in Group E. Conclusion Ocular surgery combined with systemic chemotherapy is an effective method to treat retinoblastoma. The consequences depends on the classification with higher conservative rate of eyeballs in early and middle stage (Group A, B, C), and lower rate in late stage (Group D and E).
Objective To investigate the effect of berbamine (BBM) on the proliferation and apoptosis of retinoblastoma (RB) HXO-RB44 cells and its possible mechanism in vitro.Methods RB cells in logarithmic growth phase were divided into BBM treated group and control group. RB cells in BBM treated group were cultured with different concentrations of BBM (2,4,8,16 and 32 mg/L) for 24,48 and 72 hours, respectively. The proliferation was assayed by methyl Thiazolyl tetrazolium (MTT). RB cells were cultured with different concentrations of BBM (4,8 and 16 mg/L) for 24 hours. The early apoptotic rates were detected by flow cytometry; the expression of bcl-2 and Bax were measured by enzyme-linked immunosorbent assay (ELISA) and the activity of Caspase-3 was detected by colorimetric assay.Results BBM could obviously inhibit the proliferation of RB cells in a time and dose dependent manner (24 hours: F=70.547,P＜0.01; 48 hours: F=603.438,P＜0.01; 72 hours: F=577.521,P＜0.01). The IC50 value at 24,48 and 72 hours were 25.26, 10.94 and 6.25 mg/L, respectively. Necrosis rates of control group and BBM treated group were (1.25plusmn;0.45)%, (4.10plusmn;2.95)%, (4.39plusmn;0.21)% and (10.54plusmn;4.38)% respectively; the difference between two groups was statistically significant (F=6.527,P＜0.05). Apoptotic and necrosis rates in advanced stage of control group and BBM treated group were (2.13plusmn;0.71)%, (5.45plusmn;2.31)%, (9.86plusmn;3.18)% and (11.10plusmn;1.70)%, respectively. The difference between two groups was statistically significant (F=10.845,P＜0.05). Early apoptotic rates of control group and BBM treated group were (0.51plusmn;0.26)%, (2.68plusmn;0.35)%, (5.97plusmn;0.50)% and (11.22plusmn;1.17)%, respectively. The difference between two groups was statistically significant (F=144.976,P＜0.01). In addition, BBM dose-dependently reduced bcl-2 level and increased Bax expression, causing the reduction of the bcl-2/Bax protein ratio as well as increased the Caspase-3 activity in RB cells remarkably (bcl-2: F=835.726,P＜0.01; bax: F=111.963, P＜0.01;Caspase-3:F=298.058,P＜0.01).Conclusions BBM can inhibit the proliferation and induce apoptosis or necrosis of RB cells in vitro, down regulating the expression of bcl-2, up regulating the expression of Bax. Along with increased Caspase-3 activity these may be the apoptotic mechanisms.
Objective To analyze the efficacy and safety of Intra-arterial chemotherapy (IAC) as secondly treatment in children with retinoblastoma (RB). Methods 42 eyes of 34 consecutive RB patients were enrolled in the study after intravenous chemotherapy (IVC), including 26 males and 8 females. The average age is 14.1 months. 21 cases were bilateral and 7 cases were unilateral. A total of 42 eyes of 34 patients were classified according to the International Intraocular Retinoblastoma Classification（IIRC）as group B(n=1, 2.4%), group C (n=3, 7.1%), group D (n=32, 76.2%), or group E (n=6, 14.3%). Tumor recurrence and tumor enlargement after IVC were 4 and 10 eyes respectively, accounting for 9.0% and 24.0% respectively. Sequential treatment after IVC followed by IAC were 28 eyes, accounting for 67.0%. All treatment eyes received IAC combined with laser, cryotherapy and other eye local treatment. The IAC regimen adopted the combination and alternation administration mode, by the combination of melphalan and carboplatin or the combination of melphalan and topotecan. According to the tumor changes after IAC decide whether IAC again. If tumors increased, vitreous or subretinal implants increased will be termination of IAC and enucleation. The mean follow-up time was (21.4±3.7) months after the last IAC treatment and (6.2±2.9) months after enucleation. Ocular preservation rate and complication were evaluated. Results The average IAC procedures performed on 42 eyes were (4.0±0.9). An overall ocular preservation rate of 76.2% was observed during follow-up periods due to calcification or inactivation of tumors (32 eyes), including group B (n=1, 100%), group C (n=1, 33.3%), group D (n=27, 84.4%), group E (n=3, 50%). 10 eyes were enucleated. Among them, 2 eyes of the tumor did not shrink after IAC, tumor recurrence (n=3), vitreous hemorrhage (n=3), enophthalmos (n=1), vitreous disseminated (n=1). 34 cases of children, transient eyelid oedema were 18 cases, vitreous hemorrhage and bone marrow suppression (Ⅰ-Ⅳ) were 1, 22 casese respectively. Conclusions IAC as secondly treatment is safe and effective for RB patients, however, there is still tumor recurrence. No serious ocular local and systemic complications were observed.
Objective To observe the effect of resveratrol on multidrug resistance (MDR) in human retinoblastoma cells treated. Methods RB cells in logarithmic growth phase were divided into experimental group and control group. RB cells in experimental group were cultured with different concentrations of resveratrol (6.25, 12.50, 25.00, 50.00, 100.00 mu;mol/L) for 24 and 48 hours. The proliferation (absorbance value) was assayed using methyl thiazolyl tetrazolium (MTT). RB cells were cultured with 50.00 mu;mol/L resveratrol for 48 hours. The expressions of MDR-1, cyclooxygenase-2 (COX-2)、multidrug resistance-associated protein-1 (MRP-1), glutathione-S-transferases-pi; (GST-pi;) were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The RB cells of the control group were cultured with 0.5% dimethyl sulfoxide. Results Compared with the control group, the absorbance value decreased in experimental groups (6.25, 12.50, 25.00, 50.00 mu;mol/L) in a dose dependent manner (F=4.782，P＜0.05). The difference of absorbance value between 50.00 and 100.00 mu;mol/L experimental groups was not significant (F=6.351,P＞0.05). Compared with the control group, the mRNA (t=9.170, 5.758, 4.152, 4.638) and protein (t=3.848, 5.955, 4.541, 3.514) expression levels of MDR-1, MRP1, COX-2, and GST-pi; decreased in the experimental group (P＜0.05). Conclusion Resveratrol can down-regulate the expression of MDR in RB cells.
Nowadays, one of the most challenging aspects of retinoblastoma (RB) therapy is how to control the resistant or recurrent viable vitreous seeds, for which intravenous chemotherapy appears to be ineffective. Recently, intravitreal chemotherapy offers another option to control advanced stage and vitreous seeds of RB, and may be a promising new approach to RB therapy. However, intravitreal injection for RB patients raises considerable controversy due to concerns of possible extraocular extension along the injection route, and should not replace the primary standard of care for bilateral RB or group E eyes of RB. Close follow-up and further studies are needed to determine appropriate indications, to determine the effective drugs and concentrations, to optimize RB therapy protocols and to investigate the relationship between long-term efficacy and toxicities.