Studies on the Role of Nuclear Pore Protein Translocated Promoter Region (TPR) in Carcinogenesis

13301甲第4839号博士（学術）金沢大学博士論文要旨Abstract 要約Outline 以下に掲載：Oncotarget 9(17) pp.13337-13352 2018. Impact Journals. 共著者：Firli R.P. Dewi, Takahiro Domoto, Masaharu Hazawa, Akiko Kobayashi, Takayuki Douwaki, Toshinari Minamoto, ichard W. Won

The role of nucleoporin Nup58 during mitosis

Nuclear pore complexes (NPCs) are transport channels between the nucleus and the cytoplasm. The NPCs are composed by around 30 different proteins, termed nucleoporins (Nups) and each Nup is present in multiple copies. Recently, we and others discovered that several nucleoporins play critical roles during cell division including chromosome condensation, sister chromatid cohesion, kinetochore assembly and spindle formation. Nup58 is a part of the central transport channel of the NPC, which forms a complex protein with other nucleoporins such as Nup62 and Nup54. Recently, we showed that Nup62 plays a novel role in centrosome integrity. Here, we show that Nup62 interacts with Nup58 during cell mitosis. Next, we performed RNA interference-mediated knockdown of Nup58. Currently, we are investigating nup58 depletion effect in cell cycle

The role of nucleoporin Nup58 during cell division

Nuclear pore complexes (NPCs) are transport channels between the nucleus and the cytoplasm. The NPCs are composed by around 30 different proteins, termed nucleoporins (Nups) and each Nup is present in multiple copies. Recently, we and others discovered that several nucleoporins play critical roles during cell division including chromosome condensation, sister chromatid cohesion, kinetochore assembly and spindle formation. Nup58 is a part of the central transport channel of the NPC, which forms a complex protein with other nucleoporins such as Nup62 and Nup54. Recently, we showed that Nup62 plays a novel role in centrosome integrity. Here, we show that Nup62 interacts with Nup58 during cell mitosis. Next, we performed RNA interference-mediated knockdown of Nup58. Currently, we are investigating Nup58 depletion effect in cell cycle

Enhancing the Anticancer Activity of Squamocin for Breast Cancer Treatment Using Nanodiamond Nanoparticles: An In Vivo Study

Squamocin is one of the annonaceous acetogenins produced by the Annonaceae family and displays potent anti-cancer activity against cancer cell lines. This study aimed to investigate the growth inhibition activity of squamocin coupled with nanodiamond on rats (Rattus norvegicus)-induced breast cancer. Twenty-five female R. norvegicus were divided into five groups (n = 5), including normal control (without any treatment), negative control, group treated with nanodiamond only (ND), group treated with squamocin only (SQ), and the group treated with squamocin coupled with nanodiamond (NDSQ). All of the animal models were induced for breast cancer, except for the normal control group. Breast cancer induction was performed using two doses of N-nitroso-N-methylurea (NMU) injection (50 and 30 mg/kg body weight) intraperitoneally and waited for 22 weeks until the tumor was detected to formed. Nanodiamond coupled with squamocin were administered by intraperitoneal injection (1.5 mg/kg body weight) for 5 weeks, one injection per 3 days. This study showed that the treatment with squamocin coupled with nanodiamond (NDSQ) significantly reduced the proliferation (Ki-67) and induced apoptosis (Caspase-3) of breast cancer cells, corresponding to the reduction of the thickness of the mammary ductal epithelium (p<0.001) and the lower level of CA-153 in serum. In addition, the treatment significantly reduced the malondioldehyde (MDA) and PI3KCA and increased the p53 level significantly. Altogether, in this study, we are the first to report the anti-cancer activity of squamocin in rat-induced breast cancer and the potency of nanodiamond as a carrier of squamocin to increase its anti-cancer activity

Annonacin and Squamocin Conjugation with Nanodiamond Alters Metastatic Marker Expression in Breast Cancer Cell Line

Breast cancer can perform metastasis to distant organs and cause more than 90% of malignancy-related deaths. The anti-metastasis potency of nanodiamond-conjugated annonacin and squamocin against MCF-7 cells is currently studied. First, IC50 determination of both free annonacin and squamocin to evaluate their potency as cytotoxic agents. Upon getting the IC50 value, both compounds are conjugated into nanodiamonds. Drug loading efficiencies of nanodiamond-conjugated annonacin and squamocin are 88.9% and 89.1%, respectively. Meanwhile, the ND-annonacin and ND-squamocin complex size is 150-300 nm based on SEM imaging. Subsequently, cell viability assessment of MCF-7 was performed with six cohort designs, namely, K (control cell), AN (annonacin), SQ (squamocin), NDAN (nanodiamond-conjugated annonacin), and NDSQ (nanodiamond-conjugated squamocin). Both IC50 and cell viability are assessed by MTT assay after 24 h incubation. All cohorts also underwent gene expression analysis subject to the metastasis markers CTNND1 (catenin delta 1), NOTCH4, and C-JUN. Here, the IC50 of both free annonacin (4.52 µg/ml) and squamocin (10.03 µg/ml) are more than IC50 of potent anticancer (< 4 µg/ml) for pure compounds. However, nanodiamond conjugation to both compounds can decrease cell viability better than free compounds. Compared to K, nanodiamond-conjugated annonacin and squamocin significantly decreases cell viability after 24 h incubation. Bioinformatics analysis confirmed significant pro-metastasis (C-JUN and NOTCH4) upregulation and anti-metastasis (CTNND1) downregulation in tumors compared to normal. Recent findings demonstrated that nanodiamond-conjugated annonacin can significantly upregulate CTNND1 and significantly downregulate C-JUN and NOTCH4. Even so, nanodiamond-conjugated squamocin upregulate CTNND1 but not significantly and significantly downregulate C-JUN and NOTCH4

Supplementary feed potential on histology and immune response of tilapia (Oreochromis niloticus L.) exposed to microplastics

Polyester microplastics (PS) are toxic and hazardous chemicals in the ecosystem that can induce stress. Disposing PS articles into the environment can negatively impact health of aquatic biota, including fish. This study aimed to investigate the potential of probiotics or vitamin C supplementation in improving the histological structure of organs and cytokine secretion in tilapia fish exposed to PS. Thirty-six tilapia were divided into 12 groups consisting of treatment groups (four PS concentration variations: 0, 0.1, 1, and 10 mg/L). Each treatment was given three types of feed: Commercial feed alone, commercial feed containing probiotics (200 mL/kg), and commercial feed containing vitamin C (100 mg/kg). The study had a sample size of n=3. After treatment was completed, all parameters were measured. The result showed that the addition of probiotics and vitamin C could decrease TNF-α levels and increase IFN-γ levels. Probiotics and vitamin C prevent healthy cells to be damaged by pro-inflammatory cytokines. The percentage of normal hepatocytes increased significantly in all treatment groups with the addition of probiotics or vitamin C. Furthermore, the percentage of hepatocytes with swelling and necrosis decreased significantly in treatment groups (p < 0.05). Additionally, villi height, lamina propria width, submucosa height, and the number of goblet cells all increased significantly in all treatment groups with the administration of probiotics or vitamin C (p < 0.05). Overall, both probiotics and vitamin C supplements have the potential to maintain fish health. Vitamin C exhibits a greater potential than probiotics in regulating immune responses. Meanwhile, both probiotics and vitamin C supplements have potential to inhibit damage to the hepatic and intestine structures of fish exposed to PS

THE SPATIOTEMPORAL DYNAMICS OF NUCLEOPORIN NUP58 DURING CELL DIVISION

Nuclear pore complexes (NPCs) are transport channels between the nucleus and the cytoplasm. The NPCs are composed by around 30 Nuclear pore complexes (NPCs) are transport channels between the nucleus and the cytoplasm. The NPCs are composed by around 30 different proteins, termed nucleoporins (Nups) and each Nup is present in multiple copies. Recently, we and others discovered that several nucleoporins play critical roles during cell division including centrosome integrity, kinetochore assembly, spindle formation and orientation. Nup58 is a part of the central transport channel of the NPC, which forms a complex protein with other nucleoporins such as Nup62 and Nup54. Where Nup58 localize and what their roles during cell division are still poorly understood. Here, we show that Nup58 localizes at the Nuclear pore complexes (NPCs) are transport channels between the nucleus and the cytoplasm. The NPCs are composed by around 30 different proteins, termed nucleoporins (Nups) and each Nup is present in multiple copies. Recently, we and others discovered that several nucleoporins play critical roles during cell division including centrosome integrity, kinetochore assembly, spindle formation and orientation. Nup58 is a part of the central transport channel of the NPC, which forms a complex protein with other nucleoporins such as Nup62 and Nup54. Where Nup58 localize and what their roles during cell division are still poorly understood. Here, we show that Nup58 localizes at the centrosome during prophase to metaphase, interacting with Nup62, Nup54 and also two centrosome marker protein gamma tubulin and SAS6. Depletion of Nup58 leads to monopolar spindle formation, multi-nucleic cell formation, and delayed cytokinesis. We suggest that Nup58 may regulate cytokinesis abscission. Our study gives novel insight into the role of Nup58 in cell division. centrosome during prophase to metaphase, interacting with Nup62, Nup54 and also two centrosome marker protein gamma tubulin and SAS6. Depletion of Nup58 leads to monopolar spindle formation, multi-nucleic cell formation, and delayed cytokinesis. We suggest that Nup58 may regulate cytokinesis abscission. Our study gives novel insight into the role of Nup58 in cell division. different proteins, termed nucleoporins (Nups) and each Nup is present in multiple copies. Recently, we and others discovered that several nucleoporins play critical roles during cell division including centrosome integrity, kinetochore assembly, spindle formation and orientation. Nup58 is a part of the central transport channel of the NPC, which forms a complex protein with other nucleoporins such as Nup62 and Nup54. Where Nup58 localize and what their roles during cell division are still poorly understood. Here, we show that Nup58 localizes at the centrosome during prophase to metaphase, interacting with Nup62, Nup54 and also two centrosome marker protein gamma tubulin and SAS6. Depletion of Nup58 leads to monopolar spindle formation, multi-nucleic cell formation, and delayed cytokinesis. We suggest that Nup58 may regulate cytokinesis abscission. Our study gives novel insight into the role of Nup58 in cell division

Increasing the effect of annonacin using nanodiamonds to inhibit breast cancer cells growth in rats (Rattus norvegicus)-Induced breast cancer

Background: Annonaceous acetogenins have been reported to have anti-cancer properties but low viability. In this study, we aimed to investigate the potency of nanodiamonds to be employed as a carrier of annonacin to help increase its viability and inhibit the growth of breast cancer cells. Methods: The annonacin was coupled with nanodiamond and characterized using UV-Vis spectrophotometer, FTIR, SEM, and PSA, and determined their stability and drug release. A cell growth inhibition assay and cell migration assay was performed using the breast cancer MCF7 and T747D cell lines, and in vivo analysis was performed in rats (Rattus norvegicus). MCF7 and T747D cells were treated with 12.5 μg/mL annonacin coupled with nanodiamonds for 24 and 48 h and further analyzed by MTT, cell migration, and reactive oxygen species (ROS) assays. Twenty-five female rats were divided into five groups. Breast cancer was induced using two intraperitoneal doses of N-nitroso-N-methylurea (NMU) (50 and 30 mg/kg body weight). Annonacin coupled with nanodiamonds was administered by intraperitoneal injection (17.5 mg/kg body weight) for 5 weeks, one injection per 3 days. Results: Administration of annonacin coupled with nanodiamonds significantly reduced MCF7 cell growth and reactive oxygen species (ROS) levels. The in vivo study showed that administration of annonacin coupled with nanodiamonds significantly reduced PI3KCA levels and increased p53 expression, reduced cancer antigen-15-3 (CA-15-3) levels in serum, increased caspase-3 expression, reduced Ki-67 levels, and reduced the thickness of the mammary ductal epithelium. Conclusions: Collectively, this study demonstrated the effectiveness of nanodiamonds as a carrier of annonacin to inhibit breast cancer cell growth through inhibition of the PI3K/Akt signaling pathway