• Title/Summary/Keyword: HE staining

Search Result 132, Processing Time 0.025 seconds

Effects of Tiam 1 on Invasive Capacity of Gastric Cancer Cells in vitro and Underlying Mechanisms

  • Zhu, Jin-Ming;Yu, Pei-Wu
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.1
    • /
    • pp.201-208
    • /
    • 2013
  • Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

Effects of Follicle Stimulating Hormone on ${\gamma}$-Ray Irradiated Immature Mouse Ovarian Follicles (난포성숙호르몬이 감마선 조사된 미성숙 생쥐 난포에 미치는 영향)

  • Kim, Jin-Kyu;Lee, Chang-Joo;Lee, Young-Keun;Song, Kang-Won;Yoon, Yong-Dal
    • Journal of Radiation Protection and Research
    • /
    • v.23 no.2
    • /
    • pp.89-96
    • /
    • 1998
  • To assess the radioprotective effects of follicle stimulating hormone (FSH) on ovarian follicles, 3 week-old female mice were irradiated with 8.33 Gy of ${\gamma}$-ray (group R) and followed by 5 IU ip-injection of FSH (group RF). For control groups, 5 IU of saline (group C) or 5 IU of FSH (group F) was ip-injected. Ovaries were collected 0h, 6h, 12h, 14, 2d, 4d, and 8d after irradiation or saline/FSH injection, and followed by fixation in neutral buffered formalin for routine histochemistry. Immunohistochemistry was used to assess the status of follicles and DNA fragmentation was analyzed by agarose gel electrophoresis for total DNA. Staining specific for apoptotic follicles showed high intensity at 6h and 12h in group R and RF On the other hand, staining specific for proliferating follicles showed noticeably high intensity at 8d in group R and Rf. DNA fragmentation of 185bp increased with time in all experimental groups. Especially 370bp appeared at 6h in group R, then disappeared after 1d. In case of group RF, it appeared at 12h and disappeared after 1d. From the above results, the irradiated antral follicles become completely disappeared from 4d to 8d, and then new follicles started to grow again at 8d. FSH had delaying or suppressing effects on follicular atresia after irradiation. In addition, it became clear that radiation-induced follicular atresia was mediated by granulosa cell apoptosis.

  • PDF

The Effects of Plum Extracts on the Proliferation of Human Epithelial Cell and Human Cervical Cancer Cells (자두 추출물이 인체 상피세포와 자궁경부암세포의 증식에 미치는 효과)

  • Han, Man-Deuk;Kweon, Dur-Han;Kang, Byung-Tae;Lee, Jae-Woo;Yoon, Ok-Hyun
    • Journal of the East Asian Society of Dietary Life
    • /
    • v.17 no.5
    • /
    • pp.710-718
    • /
    • 2007
  • This study was designed to investigate the effects of plum(Prunus salicina Lindl. cultivars 'Oishiwase', 'Formosa', and 'Soldam') extracts on the proliferation as well as inhibition of human epithelial cells(HaCaT), human cervical carcinoma (HeLa, SiHa, and C33A) cells, and human stomach adenocarcinoma(SNU 638) cells. Dried plum was sequentially extracted and fractionated by hexane(KC-01), chloroform(KC-02), ethyl acetate(KC-03), n-butanol(KC-04), water(KC-05), methanol(KC-6), and hot water extract(KC-07). The epithelial and cancer cells were exposed for 48 h to $50{\mu}g/mL$ of plum extract in vitro, and were then analysed by a sulforhodamin B(SRB) staining assay. The methanol extract(KCP-6) of 'Formosa' proliferated not only the HaCaT cells(147.3%), but also the cervical carcinoma C33A cells(167.8%). The ethyl acetate extract of 'Soldam'(KCJ-3) significantly reduced the proliferation rate of the HPV positive conical carcinoma cells, at 61.5% for the SiHa cells and 70.5% for the HeLa cells. In the C33A cells, which are HPV negative cervical carcinoma cells, the hexane fractions of 'Formosa'(KCP-1) and 'Oishiwase'(KCD-1) markedly suppressed proliferation activity at 20.4% and 61.7%, respectively. However, the proliferation rate of the normal epithelial cells(HaCaT cell) was not reduced the proliferation rate by KCJ-3, KCP-1, or KCD-1, There were no significant effects on proliferation of the stomach cancer cells(SNU 638) by any of the extracts or fractions of the plum cultivars. These results suggest that the anti-proliferative effects of the plum cultivars were selective to the cancer cell origin. In conclusion, we found that several plum cultivar extracts, especially, the ethyl acetate fraction of 'Soldam" and the hexane fraction of "Formosa', have anti-proliferative activity toward human cervical carcinoma cells. However, further investigation is needed to assess the molecular mechanisms that mediate the antiproliferation activities of the plum cultivars.

  • PDF

Ginsengenin derivatives synthesized from 20(R)-panaxotriol: Synthesis, characterization, and antitumor activity targeting HIF-1 pathway

  • Guo, Hong-Yan;Xing, Yue;Sun, Yu-Qiao;Liu, Can;Xu, Qian;Shang, Fan-Fan;Zhang, Run-Hui;Jin, Xue-Jun;Chen, Fener;Lee, Jung Joon;Kang, Dongzhou;Shen, Qing-Kun;Quan, Zhe-Shan
    • Journal of Ginseng Research
    • /
    • v.46 no.6
    • /
    • pp.738-749
    • /
    • 2022
  • Background: Ginseng possesses antitumor effects, and ginsenosides are considered to be one of its main active chemical components. Ginsenosides can further be hydrolyzed to generate secondary saponins, and 20(R)-panaxotriol is an important sapogenin of ginsenosides. We aimed to synthesize a new ginsengenin derivative from 20(R)-panaxotriol and investigate its antitumor activity in vivo and in vitro. Methods: Here, 20(R)-panaxotriol was selected as a precursor and was modified into its derivatives. The new products were characterized by 1H-NMR, 13C-NMR and HR-MS and evaluated by molecular docking, MTT, luciferase reporter assay, western blotting, immunofluorescent staining, colony formation assay, EdU labeling and immunofluorescence, apoptosis assay, cells migration assay, transwell assay and in vivo antitumor activity assay. Results: The derivative with the best antitumor activity was identified as 6,12-dihydroxy-4,4,8,10,14-pentamethyl-17-(2,6,6-trimethyltetrahydro-2H-pyran-2-yl)hexadecahydro-1H-cyclopenta[a]phenanthren-3-yl(tert-butoxycarbonyl)glycinate (A11). The focus of this research was on the antitumor activity of the derivatives. The efficacy of the derivative A11 (IC50 < 0.3 µM) was more than 100 times higher than that of 20(R)- panaxotriol (IC50 > 30 µM). In addition, A11 inhibited the protein expression and nuclear accumulation of the hypoxia-inducible factor HIF-1α in HeLa cells under hypoxic conditions in a dose-dependent manner. Moreover, A11 dose-dependently inhibited the proliferation, migration, and invasion of HeLa cells, while promoting their apoptosis. Notably, the inhibition by A11 was more significant than that by 20(R)-panaxotriol (p < 0.01) in vivo. Conclusion: To our knowledge, this is the first study to report the production of derivative A11 from 20(R)-panaxotriol and its superior antitumor activity compared to its precursor. Moreover, derivative A11 can be used to further study and develop novel antitumor drugs.

Renoprotective Effect of Hominis Placenta Aqua-acupuncture in Streptozotocin Induced Diabetic Rats (자하차약침(紫河車藥鍼)이 당뇨유발(糖尿誘發) 흰쥐의 신장보호기능(腎臟保護機能)에 미치는 영향(影響))

  • You, Jin-ho;Choi, Do-young;Kang, Sung-gil
    • Journal of Acupuncture Research
    • /
    • v.19 no.4
    • /
    • pp.152-166
    • /
    • 2002
  • Objective : This study was undertaken to determine whether Hominis Placenta(HP) aqua-acupuncture exerts renoprotective effect on diabetic nephropathy induced by streptozotocin(STZ) in rats. Methods : In order to study the renoprotective effect of HP aqua-acupuncture, experimental animals were divided into 5 groups and treated for 2 weeks as follows: control group was injected subcutaneously by saline aqua-acupuncture into Sinsu(BL23) in STZ induced diabetic rats, 5% HP aqua-acupuncture group was injected subcutaneously by 5% HP aqua-acupuncture into Sinsu(BL23) in STZ induced diabetic rats, 10% HP aqua-acupuncture group was injected subcutaneously by 10% HP aqua-acupuncture into Sinsu(BL23) in STZ induced diabetic rats, normal group was injected subcutaneously by saline aqua-acupuncture into Sinsu(BL23) in normal rats, and Captopril group was administrated with captopril at a dose of 50mg/kg in STZ induced diabetic rats. Results : While HP aqua-acupuncture did not reduce any body weight, index of kidney hypertrophy, the plasma glucose concentration and BUN respectively, HP aqua-acupuncture showed lowering urinary albumin excretion rate and serum creatinine as compared with the control group. Gene and protein expressions of $TGF-{\beta}1$ and fibronectin in kidney, one of the extracellular matrix proteins were investigated. There were significant differences in expression levels in HP aqua-acupuncture group as compared with the control group, and $100{\mu}g/m{\ell}$ and $500{\mu}g/m{\ell}$ of HP depressed apoptosis, showing in a dose dependent manner. In the HE staining, HP aqua-acupuncture inhibited the injury of glomerulus and proximal convoluted tubule. Conclusions : HP aqua-acupuncture showed the renoprotective effect possibly through suppressions of $TGF-{\beta}1$ and fibronectin expressions in kidney.

  • PDF

Effects of Electrical Stimulation on Wound Healing and Skin Mast Cells in Streptozotocin-Induced Diabetic Rats (스트렙토조토신 유도 당뇨 흰쥐에서 전기자극이 상처치유와 피부 비만세포에 미치는 영향)

  • Jekal, Seung-Joo;Lee, Kyung-Sun;Chung, Ok-Bong;Lee, Jae-Hyoung
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.40 no.2
    • /
    • pp.118-128
    • /
    • 2008
  • The aim of this study was to investigate the effect of electrical stimulation on healing of impaired wound and alteration of mast cells in experimental diabetic rats. Thirty male Sprague-Dawley rats were divided into three groups : incision (control), diabetes+incision (diabetes) and diabetes + incision + electrical stimulation (D/ES). Diabetes was induced in rats by streptozotocin (STZ) injection (60 mg/kg, one time) and 20 mm length incision wounds were created on the back after shaving hair. The electrical stimulation rats were treated with a current intensity of 30~50 V at 120 pps and $140{\mu}s$ for 10 days from 3 days after STZ injection. The lesion and adjacent skin tissues were fixed with 10% buffered formalin, embedded with paraffin. For wound healing analysis, hematoxylin-eosin (HE) and picrosirius red staining were performed. Mast cells (MC) were stained with toluidine blue (pH 0.5) and quantified at ${\times}200$ using a light microscope. The density of keratinocyte proliferation and microvessels in skin tissues were analyzed using a computerized image analysis system on sections immunostained with proliferative cell nuclear antigen (PCNA) and ${\alpha}$-smooth muscle actin (${\alpha}$-SMA), respectively. The results showed that the wound healing rate, collagen density and neoepidermis thickness, density of PCNA-positive cells and density of ${\alpha}$-SMA-positive vessels were significantly higher in D/ES rats than in diabetic rats. The density of MCs and degranulated MCs in D/ES rats were also significantly higher than those in diabetic rats. These findings suggest that the electrical stimulation may promote the tissue repair process by accelerating collagen production, keratinocyte proliferation and angiogenesis in the diabetic rats, and MCs are required for wound healing of skin in rats.

  • PDF

A Case of Adult Intussusception Induced by Intestinal Tuberculosis (장결핵으로 인한 성인 장중첩증 1예)

  • Kang, Hye-Sun;Kang, Ji-Young;Kang, Hyun-Hui;Kim, Hyeon-Jin;Lim, Keun-Joon;Kim, Seung-Kyoung;Lee, Sang-Haak;Moon, Hwa-Sik;Park, Jong-Kyung
    • Tuberculosis and Respiratory Diseases
    • /
    • v.69 no.3
    • /
    • pp.196-200
    • /
    • 2010
  • Intussusception primarily occurs in children and is uncommon in adults. Moreover, intussusception caused by intestinal tuberculosis is very rare. We report a case of intussusception induced by intestinal tuberculosis. A 53-year-old man presented to our hospital with complaints of cough and sputum for 2 weeks. We started anti-tuberculosis medication as the patient's sputum acid-fast staining was positive. After 4 days of treatment, the patient developed abdominal cramping pain. Imaging studies showed ileo-ileal type intussusception. The patient underwent segmental resection of the small bowel and intestinal tuberculosis was confirmed on histological examination. He recovered after surgery and was discharged on anti-tuberculosis medication.

The Effects of Pulsed Ultrasound Treatment and Cold Therapy in the Process of Acute Wound Healing (흰쥐의 급성기 창상치유에 맥동초음파와 냉 적용이 미치는 효과)

  • Kim, Tae-Yoon;Cho, Nam-Jung;Jeong, Hwa-Su;Kim, Mon-Su
    • Journal of the Korean Academy of Clinical Electrophysiology
    • /
    • v.5 no.2
    • /
    • pp.23-33
    • /
    • 2007
  • Summary of background data: At the wound from life of the human being very regarding at and a healing process the wound in the portion which is important meantime the many research is accomplished and healing process at the wound the research is small very from the physical therapy territory. Purpose: This study was performed to examine the effects of cold therapy and pulsed ultrasound treatment in the process of acute wound healing. To this end, we measured changes in the length of the wounds, and observed tissues through an optical microscope in order to evaluate the healing process of the acute wounds. Methods: We divided twenty Sprague-Dawley rats into four experimental groups of five rats each and treated them for three days after wound creation. Then we extracted tissues from the wounds on day 6th after wound creation and then took them out for hematoxylin-eosin(HE) staining. We measured changes in the length of the wounds every other day. Result: We were able to detect significant statistical differences in the cold therapy and pulsed ultrasound-treated wounds compared with control wounds. We observed tissues through an optical microscope and found the tissues in cold therapy and pulsed ultrasound-treated wounds healed well. Conclusions: Overall results indicated that the use of cold therapy and pulsed ultrasound treatment were effective in the process of acute wound healing.

  • PDF

Diagnostic and Clinical Significance of KIT(CD117) Expression in Thymic Epithelial Tumors in China

  • Song, Nan;Chen, Gang;Zhang, Peng;Liu, Ming;He, Wen-Xin;Jiang, Ge-Ning
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.6
    • /
    • pp.2745-2748
    • /
    • 2012
  • Aims: To study KIT (CD117) expression in thymic epithelial tumors in China, and investigate diagnostic and clinical significance. Material and Methods: Thymic epithelial tumors (TETs) from 102 patients (3 type A, 29 type AB, 5 type B1, 22 type B2, 29 typeB3 and 16 thymic carcinomas) were examined. Immunohistochemical staining with an antic-kit monoclonal antibody was performed on a tissue microarray. Relationships between KIT positive expression and the TET clinical characteristics (WHO histologic classification and Masaoka stage system) were analysed. Results: The KIT positive expression rate was significantly higher in thymic carcinoma (60%, 9/16) than in thymoma (8%, 7/86), a strong correlation being found with the WHO classification, but not the Masaoka tumor stage. The overall survival for patients with KIT positive lesions was significantly worse. Conclusions: KIT is a good molecule marker to differentially diagnose thymic carcinoma from thymoma, while also serving as a predictor of prognosis for TETs. Further research into KIT mutations in Chinese TETs should be conducted to assess the efficacy of targeted therapy.

Induction of Apoptosis in Glioma Cells and Upregulation of Fas Expression Using the Human Interferon-β Gene

  • Guo, Yan;Wang, Gan;Gao, Wen-Wei;Cheng, Shi-Wen;Wang, Ren;Ju, Shi-Ming;Cao, He-Li;Tian, Heng-Li
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.6
    • /
    • pp.2837-2840
    • /
    • 2012
  • We investigated whether IFN-${\beta}$ inhibits the growth of human malignant glioma and induces glioma cell apoptosis using the human IFN-${\beta}$ gene transfected into glioma cells. A eukaryonic expression vector ($pSV2IFN{\beta}$) for IFN-${\beta}$ was transfected into the glioma cell line SHG44 using liposome transfection. Stable transfection and IFN-${\beta}$ expression were confirmed using an enzyme-linked immunosorbent assay (ELISA). Cell apoptosis was also assessed by Hoechst staining and electron microscopy. In vivo experiments were used to establish a SHG44 glioma model in nude mice. Liposomes containing the human IFN-${\beta}$ gene were injected into the SHG44 glioma of nude mice to observe glioma growth and calculate tumor size. Fas expression was evaluated using immunohistochemistry. The IFN-${\beta}$ gene was successfully transfected and expressed in the SHG44 glioma cells in vitro. A significant difference in the number of apoptotic cells was observed between transfected and non-transfected cells. Glioma growth in nude mice was inhibited in vivo, with significant induction of apoptosis. Fas expression was also elevated. The IFN-${\beta}$ gene induces apoptosis in glioma cells, possibly through upregulation of Fas. The IFN-${\beta}$ gene modulation in the Fas pathway and apoptosis in glioma cells may be important for the treatment of gliomas.