siRNA’nın biyodağılımına kitozan komplekslerinin etkisi

Amaç: RNAi kanser dahil olmak üzere birçok hastalığın moleküler mekanizmasının analizinde ve gen susturulmasında hücresel proseslerin kontrolü için önemli bir araçtır. VEGF sinyali meme kanserinde siRNA taşınmasında önemli bir hedeftir. siRNA farklı hastalıklar için potansiyel bir ajan olmasına rağmen, siRNA’nın intrasellüler taşınması, terapötik olarak aktif bir moleküle dönüşmesindeki önemli engellerden biridir. Bugüne kadar birçok transfeksiyon yöntemi ve taşıyıcı sistem geliştirilmiştir. Bunlar arasında kitozan, biyouyumlu, biyoparçalanabilir olması, toksik ve immunojenik olmaması gibi özellikleri nedeniyle önemli bir gen taşıyıcısıdır. Bu çalışmanın amacı, meme kanserinde kitozan/VEGF-siRNA komplekslerinin tümör lokalizasyonunu ve biyodağılımını araştırmaktır. Yöntem: Çalışmamızda meme tümörü taşıyan sıçanlara serbest FITC-işaretli siVEGF (40 µg/sıçan) ve kitozan/ FITC-işaretli siVEGF (40 µg/sıçan) kompleksleri intravenöz olarak enjekte edildi. Bulgular: Kitozan/siVEGF komplekslerinin beyin ve kalbe biyodağılımı, serbest siVEGF ile hemen hemen benzerken, dalak, karaciğer, akciğer ve kasta biraz daha düşük ve böbrekte ise biraz daha yüksektir. Meme tümör dokusunda, kompleksler enjeksiyon sonrası 15 dakikada tümörde lokalize iken, serbest FITC-siVEGF tümör dokusunda lokalize değildir. Sonuç: Bu ön çalışmada, biz biyodağılım için VEGF siRNA taşıyıcı sistem olarak kitozanın umut verici olduğunu gösterdik

First-line treatment of patients with HER2-positive metastatic gastric and gastroesophageal junction cancer

Fluoropyrimidine+cisplatin/oxaliplatin+trastuzumab therapy is recommended for the first-line treatment of HER2-positive metastatic gastric adenocarcinoma. However, there is no comprehensive study on which platinum-based treatment should be preferred. This study aimed to compare the treatment response and survival characteristics of patients with HER2-positive metastatic gastric or gastroesophageal junction (GEJ) cancer who received fluorouracil, oxaliplatin, and leucovorin (mFOLFOX)+trastuzumab or cisplatin and fluorouracil (CF)+trastuzumab as first-line therapy. It was a multicenter, retrospective study of the Turkish Oncology Group, which included 243 patients from 21 oncology centers. There were 113 patients in the mFOLFOX+trastuzumab arm and 130 patients in the CF+trastuzumab arm. The median age was 62 years in the mFOLFOX+trastuzumab arm and 61 years in the CF+trastuzumab arm (P = 0.495). 81.4% of patients in the mFOLFOX+trastuzumab arm and 83.1% in the CF+trastuzumab arm had gastric tumor localization (P = 0.735). The median progression-free survival (PFS) was significantly higher in the mFOLFOX+trastuzumab arm (9.4 months vs. 7.3 months, P = 0.024). The median overall survival (OS) was similar in both groups (18.4 months vs. 15.1 months, P = 0.640). Maintenance trastuzumab was continued after chemotherapy in 101 patients. In this subgroup, the median OS was 23.3 months and the median PFS was 13.3 months. In conclusion, mFOLFOX+trastuzumab is similar to CF+trastuzumab in terms of the median OS, but it is more effective in terms of the median PFS in the first-line treatment of HER2-positive metastatic gastric and GEJ cancer. The choice of treatment should be made by considering the prominent toxicity findings of the chemotherapy regimens

Deneysel glomerülonefrit modelinde FK506 etkisinin immünohistokimya yöntemiyle araştırılması

Anti-Thy-1.1 glomerülonefrit, mezangial hücre proliferasyonu ve mezangial matriks artışı ile karakterize deneysel bir glomerülonefrit modelidir. Bu çalışmada deneysel anti-Thy-1.1 glomerülonefrit modelinde farklı immünohistokimyasal yöntemlerle PCNA ve a-SMA ekspresyonunu araştırdık. İmmünohistokimyasal boyama, protein ekspresyonlarını araştırmak ve ölçmek için yaygın olarak kullanılmaktadır. Double boyama yöntemi parafin kesitler ve sitolojik yaymalara uygulanabilen bir yöntemdir. Bu yöntem özel teçhizat veya teknik donanım gerektirmez. Takrolimus (FK506), otoimmün hastalıkların tedavisinde ve organ nakillerinden sonra kullanılan immünsupressif bir ilaçtır. Bu çalışmanın amacı, deneysel glomerülonefrit modelinde Prolifere Hücre Nükleer Antijen (PCNA) ve a-düz kas aktin "a-Smooth Muscle Actin" (a-SMA) immünekpresyonlarını, Streptavidin-Biotin /Alkalen Fosfataz kompleks (Str.ABC/AP), Streptavidin-Biotin/Horseradish Peroksidaz kompleks (Str.ABC/HRP) ve double immünohistokimya (Str.ABC/HRP ve Str.ABC/AP) boyama metotları ile araştırmaktır. Ayrıca deneysel glomerülonefrit modelinde takrolimus etkisini de gözden geçirmektir. Yirmi Wistar Albino sıçan bu çalışmada 5'erli dört gruba ayrılmıştır. Grup I Kontrol (K): Dört hafta 0,1ml/100g serum fizyolojik (SF) intravenöz uygulanmıştır; grup II Glomerülonefrit (GN): 0.gün anti-Thy 1.1 antikor (0,25µg/100g) intravenöz uygulanmıştır; grup III Glomerülonefrit+FK506 (GFK): önce anti-Thy 1.1 antikor (0,25µg/100g) 0.günde, sonra FK506 (1mg/kg) iki hafta süresince uygulanmıştır. Grup IV Kontrol+FK506 (KFK): önce iki hafta SF (0,1ml/100g), daha sonra iki hafta boyunca FK506 (1mg/kg ) uygulanmıştır. Çalışma sonunda serumda kreatinin, idrarda kreatin, kreatin klirensi ve proteinüri bakılmıştır. Böbrek dokusunda; ışık mikroskobunda, glomerüler ve tubulointerstisyel alanda, PCNA ve a-SMA immünekspresyonu Str.ABC/HRP, Str.ABC/AP ve double immünohistokimya ile boyanan lamlarda değerlendirilmiştir. Çalışma sonunda; kontrol grubuna göre, glomerülonefrit grubunda glomerüler hücre proliferasyonu ve tubulointerstisyel hasar anlamlı ölçüde artmıştır. Tüm gruplarda uyguladığımız immünohistokimyasal metotlar ile a-SMA ekspresyonu birbirine yakın bulunmuştur. a-SMA nin tubulointerstisyal alanda immünekspresyonu Str.ABC/AP ve Str.ABC/HRP boyama yöntemlerinde GFK grubunda GN grubuna göre anlamlı derecede azalmıştır. Tüm immünohistokimya yöntemlerinde tubulointerstisyel alan ve glomerüllerde PCNA immünekspresyonu K grubuna göre GN grubunda, KFK grubuna göre GFK'de arttığı bulunmuştur. Sonuçta double immünohistokimya boyama yönteminin, aynı bölgede iki farklı antijenin araştırılmasında kullanılabilecek güvenilir ve basit bir yöntem olduğu gösterilmiştir. EFFECT OF FK506 IN EXPERIMENTAL MESANGIAL PROLIFERATIVE GLOMERULONEPHRITIS BY IMMUNOSTAINING METHODS SUMMARY The anti-Thy-1.1 model is a rat model of mesangial proliferative glomerulonephritis characterized by mesangial cell proliferation and accumulation of mesangial matrix expansion. In present study we investigate the expression of a-smooth muscle actin (a-SMA) and proliferative cell nuclear antigen (PCNA) in renal cortex in the rat model of mesangial proliferative glomerulonephritis induced with anti-Thy-1.1 antibody by different immunohistochemical methods. Immunostaining is used widely for detection and measurement of protein expression. The double staining method described may have wide application since it can be used both for paraffin sections and cell smears. It does not require specialized technique or aparatus. Tacrolimus (FK506) is an immunsuppressive drug used for the treatment of autoimmune disease and after organ transplantations. The aim of our study to evaluate the immunexpression PCNA and a-SMA in anti-Thy-1.1 diffuse experimental glomerulonephritis rat model by Str.ABC/HRP, StrABC/AP and double (Str.ABC/HRP and Str.ABC/AP) immunohistochemistry methods. Also we want to investigate tacrolimus effect in this experimental glomerulonephritis model. Twenty male Wistar Albino rats were divided into four groups of fifth: Group I Control (C): 5 treated 0,1ml /100g normal saline intravenously (IV) for four weeks; group II glomerulonephritis (GN): 5 treated with anti-Thy 1.1 (0,25µg/100g) IV at 0 day; group III Glomerulonephritis+FK506 (GFK): 5 treated with Anti-Thy-1.1 (0,25µg/100g) IV at 0 day then FK506 (1mg/kg) for two weeks. Group IV Control+FK506 (CFK): 5 treated with normal saline (0.1mg/100mg) both for two weeks then FK506 (1mg/kg) for two weeks. Serum creatinine, urinary creatinine, creatinine clearance and proteinuria were performed at the end of the study period. Renal tissue were assessed for light microscopic findings glomerul and tubulointerstitial injury and PCNA, a-SMA expression were semiquantatitively scored on glomeruli and tubulointerstitium by Str.ABC/HRP, Str.ABC/AP and double immunohistochemical methods staining slides. At the end of the study period morphological changes including tubulointerstitial injury and glomerular cell proliferations were significantly increased in glomeulonephritis group compared to control group. With in immunohistochemical methods, glomerular immunoexpression of a-SMA was similar in all groups. Tubulointestitial immunoexpression of a-SMA was significantly statical decreased in Str.ABC/AP and Str.ABC/HRP in GFK compared to GN group. Immunoexpression of PCNA in all immunohistochemical methods on tubulointerstitial and glomeruli was significant increased in GN group compared to C, and GFK group compared to CFK. In conclusion double ımmunostaing was the method to detect two antigen in same site. That method was simple and reliable

Doksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podositler üzerine etkisi : ışık Ve elektron mikroskobu çalışması

Doksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podositler üzerine etkisi : ışık Ve elektron mikroskobu çalışması Amaç: Nefrotik sendrom, böbrek yetmezliğine ilerleyebilen ve podosit hasarı oluşturan bir hastalıktır. Bir vazoaktif peptid olan apelin, etkisini apelin reseptörüne (APJ) bağlanarak gösterir. Glomerüler arteriollerin endotelyal ve vasküler düz kas hücrelerinde bulunan apelinin, böbrek hemodinamiğini düzenleyerek pre ve post mikrodamarlanma üzerinde etkisinin olduğu görülmüştür. Çalışmamızın amacı, doksorubisin ile oluşturulan nefrotik sendrom modelinde, apelinin proteinüri ve podosit hasarı üzerindeki etkisini, ışık ve elektron mikroskobu seviyesinde incelemek, bununla birlikte APJ’nin varlığını araştırmaktır. Gereç ve Yöntem: Çalışmada yetişkin erkek Sprague-Dawley sıçanlar kullanıldı. Sıçanlar randomize olarak (n=6) Kontrol (K; 1ml/kg i.p. serum fizyolojik (SF)); Apelin Kontrol (APK; SF ve 50 mcg/kg/gün Apelin-13 i.p.); Nefrotik Sendrom (NS; i.p. 10 mg/kg doksorubisin (DOX)) ve Nefrotik Sendrom+Apelin (NSAP; i.p. 10 mg/kg DOX+50 mcg/kg/gün Apelin-13) gruplarına ayrıldı. Yirmibirinci günün bitiminde idrar örnekleri toplandı; biyokimyasal, western blot, ışık ve elektron mikroskobu incelemeleri için böbrek dokuları alındı. Bulgular: Nefrotik sendrom grubunda proteinürinin K grubuna göre anlamlı olarak arttığı görüldü. Oksidatif stres belirteçleri olan MDA, NO, Lusigenin ve Luminolün NS grubunda anlamlı şekilde arttığı, NSAP grubunda ise azaldığı tespit edildi. GSH’ın ise NS grubunda anlamlı olarak azaldığı, NSAP grubunda arttğı izlendi. Işık mikroskobu incelemelerinde NS grubunda glomerüllerde mezangial matriks artışı ve tübüler hasar görüldü. NSAP grubunda ise minimal hasar gerilemesi gözlendi. Yine NS grubunda TGF-β immünekspresyonunda artış, NSAP grubunda minimal azalma gözlendi. Nefrin ve Apelin reseptör ekspresyonunda ise NS grubunda azalma görüldü. NS grubunda apoptotik hücre sayısı anlamlı artış gösterdi. Elektron mikroskobu incelemelerinde NS ve NSAP grubunda podosit ayaksı çıkıntılarında düzleşme görüldü. Tüm gruplarda podositlerde immün-altın işaretleme ile APJ varlığı gösterildi. Sonuçlar: Doksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podosit hasarı üzerinde iyileştirici etkisinin minimal olduğu gözlendi. Anahtar Sözcükler: glomerül, nefrotik sendrom, podosit, apelin, APJ. SUMMARY Histology and Embryology Department Purpose: Nephrotic syndrom is a disease which may progress to podocyte degeneration and renal failure. Apelin, is a vasoactive peptide, shows its effect by binding the apelin receptor (APJ). Apelin, present in endotelial and vascular smooth muscle cells of glomerular arteriols has effects on the pre- and post- microvascularization through regulating renal hemodynamics. The purpose of our study is to determine the effects of apelin on the proteinuria and podocyte damage at light and electron microscopic levels, also to investigate the existence of APJ in the podocytes. Materials and Methods: Male Sprague-Dawley rats were used in the study. Rats were randomly divided into 4 groups (n=6) as Control (C; i.p. 1ml/kg physiological saline solution (PS)); Apelin Control (APC; PS and 50 mcg/kg/day Apelin-13 i.p.); Nephrotic Syndrome (NS; i.p. 10 mg/kg doxorubicin (DOX)) and Nephrotic Syndrome+Apelin (NSAP; i.p. 10 mg/kg DOX+50 mcg/kg/day Apelin-13). Urine samples and kidney tissues were collected at the end of 21st day for biochemical and western blot analyses, light and electron microscopic investigations. Results: In NS group, the significant increase in proteinuira was observed when compared with control group. The markers for the oxidative stress MDA, NO, Lucigenin ve Luminol, were increased significantly in NS while were decreased in the NSAP group. A significant decrease in GSH levels in NS group while an increase in the NSAP group was observe. In light microscopy, NS group revealed mesangial matrix accumulation in glomerules besides tubular damage. The minimal regression in the damage were seen in NSAP group. Immunoexpression of TGF- β were increased in NS group while there was a decrease in NSAP group. In NS group, expression of nephrin and APJ were decreased and the apoptotic cell number was increased significanty. In electron microscopic investigations, effacement of podocyte foot process were observed in NS and NSAP groups. In all groups, immunogold labelling of APJ were shown. Conclusion: Apelin has limited therapeutic effects on podocyte damage in experimental model of nephrotic syndrome induced by doxorubucin. Keywords: glomerulus, nephrotic syndrome, podocyte, apelin, APJ

Doksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podositler üzerine etkisi : ışık Ve elektron mikroskobu çalışması

ÖZETDoksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podositler üzerine etkisi : ışık Ve elektron mikroskobu çalışmasıAmaç: Nefrotik sendrom, böbrek yetmezliğine ilerleyebilen ve podosit hasarı oluşturan bir hastalıktır. Bir vazoaktif peptid olan apelin, etkisini apelin reseptörüne (APJ) bağlanarak gösterir. Glomerüler arteriollerin endotelyal ve vasküler düz kas hücrelerinde bulunan apelinin, böbrek hemodinamiğini düzenleyerek pre ve post mikrodamarlanma üzerinde etkisinin olduğu görülmüştür. Çalışmamızın amacı, doksorubisin ile oluşturulan nefrotik sendrom modelinde, apelinin proteinüri ve podosit hasarı üzerindeki etkisini, ışık ve elektron mikroskobu seviyesinde incelemek, bununla birlikte APJ’nin varlığını araştırmaktır.Gereç ve Yöntem: Çalışmada yetişkin erkek Sprague-Dawley sıçanlar kullanıldı. Sıçanlar randomize olarak (n=6) Kontrol (K; 1ml/kg i.p. serum fizyolojik (SF)); Apelin Kontrol (APK; SF ve 50 mcg/kg/gün Apelin-13 i.p.); Nefrotik Sendrom (NS; i.p. 10 mg/kg doksorubisin (DOX)) ve Nefrotik Sendrom+Apelin (NSAP; i.p. 10 mg/kg DOX+50 mcg/kg/gün Apelin-13) gruplarına ayrıldı. Yirmibirinci günün bitiminde idrar örnekleri toplandı; biyokimyasal, western blot, ışık ve elektron mikroskobu incelemeleri için böbrek dokuları alındı.Bulgular: Nefrotik sendrom grubunda proteinürinin K grubuna göre anlamlı olarak arttığı görüldü. Oksidatif stres belirteçleri olan MDA, NO, Lusigenin ve Luminolün NS grubunda anlamlı şekilde arttığı, NSAP grubunda ise azaldığı tespit edildi. GSH’ın ise NS grubunda anlamlı olarak azaldığı, NSAP grubunda arttğı izlendi. Işık mikroskobu incelemelerinde NS grubunda glomerüllerde mezangial matriks artışı ve tübüler hasar görüldü. NSAP grubunda ise minimal hasar gerilemesi gözlendi. Yine NS grubunda TGF-β immünekspresyonunda artış, NSAP grubunda minimal azalma gözlendi. Nefrin ve Apelin reseptör ekspresyonunda ise NS grubunda azalma görüldü. NS grubunda apoptotik hücre sayısı anlamlı artış gösterdi. Elektron mikroskobu incelemelerinde NS ve NSAP grubunda podosit ayaksı çıkıntılarında düzleşme görüldü. Tüm gruplarda podositlerde immün-altın işaretleme ile APJ varlığı gösterildi.Sonuçlar: Doksorubisin ile oluşturulan deneysel nefrotik sendrom modelinde apelinin podosit hasarı üzerinde iyileştirici etkisinin minimal olduğu gözlendi.Anahtar Sözcükler: glomerül, nefrotik sendrom, podosit, apelin, APJ.SUMMARYHistology and Embryology DepartmentPurpose: Nephrotic syndrom is a disease which may progress to podocyte degeneration and renal failure. Apelin, is a vasoactive peptide, shows its effect by binding the apelin receptor (APJ). Apelin, present in endotelial and vascular smooth muscle cells of glomerular arteriols has effects on the pre- and post- microvascularization through regulating renal hemodynamics. The purpose of our study is to determine the effects of apelin on the proteinuria and podocyte damage at light and electron microscopic levels, also to investigate the existence of APJ in the podocytes.Materials and Methods: Male Sprague-Dawley rats were used in the study. Rats were randomly divided into 4 groups (n=6) as Control (C; i.p. 1ml/kg physiological saline solution (PS)); Apelin Control (APC; PS and 50 mcg/kg/day Apelin-13 i.p.); Nephrotic Syndrome (NS; i.p. 10 mg/kg doxorubicin (DOX)) and Nephrotic Syndrome+Apelin (NSAP; i.p. 10 mg/kg DOX+50 mcg/kg/day Apelin-13). Urine samples and kidney tissues were collected at the end of 21st day for biochemical and western blot analyses, light and electron microscopic investigations.Results: In NS group, the significant increase in proteinuira was observed when compared with control group. The markers for the oxidative stress MDA, NO, Lucigenin ve Luminol, were increased significantly in NS while were decreased in the NSAP group. A significant decrease in GSH levels in NS group while an increase in the NSAP group was observe. In light microscopy, NS group revealed mesangial matrix accumulation in glomerules besides tubular damage. The minimal regression in the damage were seen in NSAP group. Immunoexpression of TGF- β were increased in NS group while there was a decrease in NSAP group. In NS group, expression of nephrin and APJ were decreased and the apoptotic cell number was increased significanty. In electron microscopic investigations, effacement of podocyte foot process were observed in NS and NSAP groups. In all groups, immunogold labelling of APJ were shown. Conclusion: Apelin has limited therapeutic effects on podocyte damage in experimental model of nephrotic syndrome induced by doxorubucin. Keywords: glomerulus, nephrotic syndrome, podocyte, apelin, APJ

BRAFV600E Immunohistochemistry in Papillary Thyroid Carcinomas: Relationship Between Clinical and Morphological Parameters

Objective: To investigate the association of the BRAFV600E mutation with papillary thyroid carcinoma using clinical, morphological and prognostic parameters. We also intend to assess the utility of the BRAFV600E immunohistochemistry and compare it with BRAF polymerase chain reaction (RT-PCR). Material and Method: We applied BRAFV600E immunohistochemistry in a cohort of 107 papillary carcinomas, 19 adenomas and 13 normal thyroid tissues that was chosen retrospectively between 2011 and 2015. Statistical analysis was based on semiquantitative immunohistochemistry findings. We also applied BRAF RT-PCR in a subgroup of 14 papillary carcinomas, 13 metastatic lymph nodes and 4 adenomas that was chosen randomly. Results: In regard to the comparison of BRAFV600E immunohistochemistry and BRAF RT-PCR, a 3+ nuclear and cytoplasmic immunoexpression was considered 'positive The BRAFV600E mutation was most frequently observed in classic variant cases. No mutation was detected in follicular variant cases. The mutational status of the primary tumour and the lymph node metastasis was consistent. A significant relationship of the BRAFV600E mutation was found with prognostic factors such as higher pT stage, classic variant, lymphatic invasion, perineural invasion, lower mitotic index, lack of tumour capsule, intrathyroidal spread and extrathyroidal extension. Conclusion: Immunohistochemistry, using the VE1 clone, is a reliable technique for detection of the BRAFV600E mutation. Our results with immunohistochemistry are consistent with a previous effort. In our study, despite the correlation between some pathological prognostic parameters and the BRAFV600E mutation; poor prognosis was found to be irrelevant overall. Morphological parameters seem to be keener than the BRAFV600E mutation. Nevertheless, different series display different results, possibly due to environmental factors. Considering this and the proven success of targeted therapies against the BRAFV600E mutation a thorough assessment would be important

Histomorphological Changes of Apelin Treatment in Renal Tissue in Doxorubicin-induced Nephrotic Syndrome Model

Objective: Nephrotic syndrome (NS) may result in renal failure. Apelin (AP), a vasoactive peptide, demonstrates its effect by binding to the AP receptor. AP, present in the endothelial and vascular smooth muscle cells of glomerular arterioles, affects the pre- and post-microvascularization by regulating renal hemodynamics. This study aimed to determine the histomorphological changes of AP treatment in a doxorubicin (DOX)-induced NS model. Methods: Male Sprague Dawley rats were used. Rats were randomly divided into four groups (n=6): control [physiological saline (PS) solution intraperitoneal (i.p.)]; AP+PS (PS and 50 mcg/kg/day AP-13 i.p.); NS [10 mg/kg DOX i.p.] and NS+AP (NSAP; 10 mg/kg DOX+50 mcg/kg/day AP-13 i.p.). Renal tissues were collected on the 22nd day for light microscopic investigations. Results: Light microscopic investigations showed that the NS group revealed adhesions between the tuft and Bowman's capsule, mesangial matrix accumulation in the glomeruli, and tubular damage with dilatation and cast accumulation. In the NSAP group, minimal regression in the glomerular and tubular damage was observed. Conclusion: The histomorphological changes of AP treatment in a DOX-induced NS model demonstrated a limited therapeutic effect on glomerular and tubular damage in renal tissues

Investigation of the Therapeutic Efficacy of Codelivery of psiRNA-Vascular Endothelial Growth Factor and pIL-4 into Chitosan Nanoparticles in the Breast Tumor Model

Angiogenesis has been known to increase tumor growth and for its metastatic potential in human tumors. Vascular endothelial growth factor (VEGF) plays an important role in tumor angiogenesis and is a promising therapeutic target for breast cancer. VEGF is an essential target for RNAi-based gene therapy of breast cancer. Interleukin-4 (IL-4) may act as an anti-angiogenic molecule that inhibits tumor growth and migration in rats. The purpose of the present study was to improve therapeutic efficacy in breast cancer with the codelivery of siRNA-expressing plasmid targeting VEGF and IL-4-expressing plasmid encapsulating into chitosan nanoparticles (NPs). The codelivery of psiVEGF and pIL-4 plasmids greatly enhanced in vitro and in vivo gene-silencing efficiency. For the in vitro study, when psiVEGF and pIL-4 into chitosan NPs were combined (81%), the gene-silencing effect was higher than psiVEGF and pIL-4 NPs alone. The in vivo study breast tumor model demonstrated that the administration of coencapsulation of psiVEGF and pIL-4 into chitosan NPs caused an additive effect on breast tumor growth inhibition (97%), compared with containing NPs psiVEGF or pIL-4 alone. These results indicate that chitosan NPs can be effectively used for the codelivery of pIL-4 and siVEGF-expressing plasmid in a combination therapy against breast cancer. (c) 2013 Wiley Periodicals, Inc

Local Delivery of Chitosan/VEGF siRNA Nanoplexes Reduces Angiogenesis and Growth of Breast Cancer In Vivo

Vascular endothelial growth factor (VEGF) is the important angiogenic factor associated with tumor growth and metastasis in a wide variety of solid tumors. The aim of this study is to investigate the tumor suppressive effect of chitosan/small interfering RNA (siRNA)-VEGF nanoplexes in the rat breast cancer model. Chitosan/siRNA nanoplexes (siVEGF-A, siVEGFR-1, siVEGFR-2) and NRP-1 were prepared in a 15 to1 ratio and injected (intratumorally) into the breast-tumor-bearing Sprague-Dawley rats. Tumor volumes were measured during 21 days. To investigate the effect of chitosan/siRNA nanoplexes on VEGF expression in tumors, VEGF was analyzed with immunohistochemistry and western blotting. The mRNA levels of VEGF in tumor samples were determined with real-time PCR (RT-PCR). After siRNA treatment, a marked reduction in tumor volumes was measured in complex-injected rats (97%). Free siRNA injection showed lower tumor inhibition. Reduction of VEGF protein was also shown with western blotting and immunohistochemistry. Similar results were obtained with RT-PCR also. These results indicate that the chitosan/siRNA targeting to VEGF nanoplexes have a remarkably suppressive effect on VEGF expression and tumor volume in breast cancer model of rats