Sex-dependent liver cancer xenograft models for predicting clinical data in the evaluation of anticancer drugs

Abstract Background The incidence and mortality of liver cancer show a great difference between the sexes. We established sex-dependent liver cancer xenograft models and investigated whether such sex-dependent models could be used to simultaneously evaluate the therapeutic and adverse effects of anticancer drugs for drug screening. Results In the in-vitro test, the cytotoxicity of anticancer drugs (cisplatin, 5-fluorouracil, and doxorubicin) was compared between male- and female-derived liver cancer cell lines. Cisplatin and 5-fluorouracil exhibited cytotoxicity without sex-difference, but doxorubicin showed dose-dependently significant cytotoxicity only in male-derived cells. Our results showed a strong correlation between preclinical and clinical data with the use of sex-dependent liver cancer xenograft models. Moreover, the male-derived Hep3B-derived xenograft model was more sensitive than the female-derived SNU-387-derived xenograft model against doxorubicin treatment. Doxorubicin showed more severe cardiotoxicity in the male xenograft model than in the female model. We investigated the occurrence frequency of doxorubicin-related cardiotoxicity using data obtained from the Korea Institute of Drug Safety & Risk Management Database, but no significant difference was observed between the sexes. Conclusions Our results suggest that sex-dependent xenograft models are useful tools for evaluating the therapeutic and adverse effects of anticancer drugs, because sex is an important consideration in drug development.

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In Vitro and In Vivo Targeting and Therapy of an Antibody-Drug Conjugate (IMMU-110) in B-Cell Malignancies.

Blood ◽

10.1182/blood.v104.11.3287.3287 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3287-3287

Author(s):

Puja Sapra ◽

Rhona Stein ◽

Jennifer Pickett ◽

Serengulam V. Govindan ◽

Thomas M. Cardillo ◽

...

Keyword(s):

B Cell ◽

Cell Lines ◽

Specific Antibody ◽

Xenograft Model ◽

B Cell Lymphoma ◽

Scid Mice ◽

Significant Difference ◽

Xenograft Models

Abstract IMMU-110 is a drug immunoconjugate comprised of doxorubicin (DOX) conjugated to the humanized anti-CD74 monoclonal antibody (mAb), hLL1, at a DOX:mAb (mol/mol) ratio of 8:1. CD74 is a rapidly internalizing type-II transmembrane chaperone molecule associated with HLA-DR, and has high expression on human non-Hodgkin’s lymphoma (NHL) and multiple myeloma (MM) clinical specimens and cell lines. Here, we investigated the in vitro and in vivo efficacy of IMMU-110 in xenograft models of human NHL (Raji, Daudi) and MM (MC/CAR). In vitro cell binding of IMMU-110 with the CD74-positive cells was significantly higher than that of a non-specific isotype-matched mAb-DOX conjugate (DOX conjugated to a mAb against epithelial glycoprotein-1; DOX-hRS7), and was similar to that of naked hLL1. Both IMMU-110 and naked hLL1 bound CD74 with subnanomolar affinity. The in vitro cytotoxicity of IMMU-110 was significantly higher than non-specific antibody-DOX conjugate, DOX-hRS7, and was similar to free DOX in MC/CAR, Raji or Daudi human Burkitt’s lymphoma cells. In CD74-negative cell lines, IMMU-110 was significantly less toxic than free DOX, having similar cytotoxicity to DOX-hRS7. In vivo, IMMU-110 displayed a pharmacokinetic and biodistribution profile almost identical to that of hLL1 mAb. Both hLL1 mAb and IMMU-110 had a biphasic clearance from the circulation; the α and β half-life (t1/2) of IMMU-110 were 4.6 h and 157.9 h, respectively, and those of hLL1 were 5.4 h and 151.5 h, respectively. In biodistribution studies, no significant difference was observed between IMMU-110 and naked hLL1 with regards to normal tissue uptake. Neither IMMU-110 nor naked hLL1 mAb had a significant association with any normal body tissue. In therapy experiments, a single i.v. protein dose of 350 μg IMMU-110, injected 5 days after implantation of MC/CAR cells in SCID mice, resulted in curing 70% of the animals. Similar cure rates were observed when treatment with IMMU-110 was given 10 days after transplantation of MC/CAR cells. In the Raji xenograft model, 100% of animals were cured with a single protein dose of 120 μg IMMU-110, injected 5 days after implantation of cells. In survival studies, the efficacy of IMMU-110 was significantly better than naked hLL1, the combination of naked hLL1 and free DOX, or of a non-specific antibody-DOX conjugate, DOX-hRS7. In a tolerability study in SCID mice, no toxic effect of IMMU-110 was observed even at the highest dose tested (2.5 mg /mouse). In conclusion, treatment of B-cell lymphoma and myeloma xenograft models with single injections of IMMU-110 resulted in high levels of response and long-term survivors. IMMU-110 is being further developed as a potential therapeutic for the treatment of CD74-positive tumors.

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APTO-253 Induces KLF4 to Promote Potent in Vitro Pro-Apoptotic Activity in Hematologic Cancer Cell Lines and Antitumor Efficacy As a Single Agent and in Combination with Azacitidine in Animal Models of Acute Myelogenous Leukemia (AML)

Blood ◽

10.1182/blood.v124.21.4813.4813 ◽

2014 ◽

Vol 124 (21) ◽

pp. 4813-4813 ◽

Cited By ~ 1

Author(s):

William G Rice ◽

Avanish Vellanki ◽

Yoon Lee ◽

Jeff Lightfoot ◽

Robert Peralta ◽

...

Keyword(s):

Bone Marrow ◽

High Risk ◽

Animal Models ◽

Antitumor Activity ◽

Single Agent ◽

Xenograft Model ◽

Myelogenous Leukemia ◽

Acute Myelogenous ◽

Xenograft Models

Abstract APTO-253, a small molecule that mediates anticancer activity through induction of the Krüppel-like factor 4 (KLF4) tumor suppressor, is being developed clinically for the treatment of acute myelogenous leukemia (AML) and high risk myelodysplastic syndromes (MDS). APTO-253 was well tolerated in a Phase I study in patients with solid tumors using a dosing schedule of days 1, 2, 15, 16 of a 28 day cycle (2T-12B-2T-12B), but recent scientific observations guided APTO-253 toward AML and high risk MDS. Indeed, suppression of KLF4 was reported as a key driver in the leukemogenesis of AML and subsets of other hematologic diseases. The vast majority (~90%) of patients with AML aberrantly express the transcription factor CDX2 in human bone marrow stem and progenitor cells (HSPC) (Scholl et al., J Clin Invest. 2007, 117(4):1037-48). The CDX2 protein binds to CDX2 consensus sequences within the KLF4 promoter, thereby suppressing KLF4 expression in HSPC (Faber et al., J Clin Invest. 2013, 123(1):299-314). Based on these observations, the anticancer activity of APTO-253 was examined in AML and other hematological cancers. APTO-253 showed potent antiproliferative activity in vitro against a panel of blood cancer cell lines, with ηM IC50values in AML (6.9 - 305 ηM), acute lymphoblastic leukemia and chronic myeloid leukemia (39 – 250 ηM), non-Hodgkin’s lymphoma (11 – 190 ηM) and multiple myeloma (72 – 180 ηM). To explore in vivo efficacy, dose scheduling studies were initially conducted in the H226 xenograft model in mice. In the H226 model, APTO-253 showed improved antitumor activity when administered for two consecutive days followed by a five day break from dosing (2T-5B) each week, i.e. on days 1,2, 8,9, 15,16, 22,23, compared to the 2T-12B-2T-12B schedule. The 2T-5B schedule was used to evaluate antitumor activity of APTO-253 in several AML xenograft models in mice. In Kasumi-1 AML and KG-1 AML xenograft models, APTO-253 showed significant antitumor activity (p = 0.028 and p=0.0004, respectively) as a single agent when administered using the 2T-5B schedule each week for four weeks compared to control animals. Mice treated with APTO-253 had no overt toxicity based on clinical observations and body weight measurements. Mice bearing HL-60 AML xenograft tumors were treated with APTO-253 for one day or two consecutive days per week for three weeks, either as a single agent or combined with azacitidine, or with azacitidine alone twice per week (on days 1,4, 8, 11, 15 and 18). APTO-253 as a single agent inhibited growth of HL-60 tumors to approximately the same extent as azacitidine. Furthermore, both once weekly and twice weekly dosing of APTO-253 in combination with azacitidine resulted in significantly enhanced antitumor activity relative to either single agent alone (p = 0.0002 and p = 0.0006 for 1X and 2X weekly APTO-253 treatment, respectively, compared to control). Likewise, using a THP-1 AML xenograft model, APTO-253 administered as a single agent using the 2T-5B per week schedule showed significant efficacy, similar to that of azacitidine, while the combination of APTO-253 and azacitidine demonstrated greatly improved antitumor effects relative to either drug alone. APTO-253 was effective and well tolerated as a single agent or in combination with azacitidine in multiple AML xenograft models, plus APTO-253 does not cause bone marrow suppression in animal models or humans. Taken together, our results indicate that APTO-253 may serve as a targeted agent for single agent use and may provide enhanced efficacy to standard of care chemotherapeutics for AML and other hematological malignancies. Disclosures Rice: Lorus Therapeutics Inc.: Employment. Vellanki:Lorus Therapeutics Inc.: Employment. Lee:Lorus Therapeutics Inc.: Employment. Lightfoot:Lorus Therapeutics Inc.: Employment. Peralta:Lorus Therapeutics Inc.: Employment. Jamerlan:Lorus Therapeutics Inc.: Employment. Jin:Lorus Therapeutics Inc.: Employment. Lum:Lorus Therapeutics Inc.: Employment. Cheng:Lorus Therapeutics Inc.: Employment.

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VEGF/Flk1 Mechanism is Involved in Roxarsone Promotion of Rat Endothelial Cell Growth and B16F10 Xenograft Tumor Angiogenesis

Scientific Reports ◽

10.1038/s41598-019-53870-3 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Shihao Chen ◽

Jinge Xu ◽

Qianhan Wei ◽

Zeting Zhao ◽

Xin Chen ◽

...

Keyword(s):

Cell Growth ◽

Cell Viability ◽

Vascular Endothelial Cells ◽

Growth Promotion ◽

Xenograft Model ◽

Feed Additive ◽

Vascular Endothelial ◽

Mouse Xenograft Model ◽

Significant Difference

AbstractThe potential angiogenic effect of roxarsone, a feed additive widely used to promote animal growth worldwide, was demonstrated recently. We explored the mechanism of vascular endothelial growth factor (VEGF) and its receptor (VEGFR) in roxarsone promotion of rat vascular endothelial cells (ECs) and B16F10 mouse xenografts. ECs were treated with 0.1–50 μM roxarsone or with roxarsone plus 10 ng/mL VEGF, VEGFR1 (Flt1), or VEGFR2 (Flk1) antibodies for 12–48 h to examine their role in cell growth promotion. Small interfering RNA (siRNA) targeting Vegf, Flt1, and Flk1 were transfected in the ECs, and we measured the expression level, cell proliferation, migration, and tube formation ability. The siRNA targeting Vegf or Flk1 were injected intratumorally in the B16F10 xenografts of mice that received 25 mg/kg roxarsone orally. Cell viability and VEGF expression following roxarsone treatment were significantly higher than that of the control (P < 0.05), peaking following treatment with 1.0 μM roxarsone. Compared to roxarsone alone, the VEGF antibody decreased cell promotion by roxarsone (P < 0.05), and the Flk1 antibody greatly reduced cell viability compared to the Flt1 antibody (P < 0.01). Roxarsone and Flk1 antibody co-treatment increased supernatant VEGF significantly, while cellular VEGF was obviously decreased (P < 0.01), whereas there was no significant difference following Flt1 antibody blockade. The siRNA against Vegf or Flk1 significantly attenuated the roxarsone promotion effects on EC proliferation, migration, and tube-like formation (P < 0.01), whereas the siRNA against Flt1 effected no obvious differences. Furthermore, the RNA interference significantly weakened the roxarsone-induced increase in xenograft weight and volume, and VEGF and Flk1 expression. Roxarsone promotion of rat EC growth, migration, and tube-like formation in vitro and of B16F10 mouse xenograft model tumor growth and angiogenesis involves a VEGF/Flk1 mechanism.

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Marked Therapeutic Efficacy of a Novel Polyethyleneglycol-SN38 Conjugate, EZN-2208, in Xenograft Models of Non-Hodgkin’s Lymphoma.

Blood ◽

10.1182/blood.v110.11.1397.1397 ◽

2007 ◽

Vol 110 (11) ◽

pp. 1397-1397

Author(s):

Puja Sapra ◽

Mary Mehlig ◽

Jennifer Malaby ◽

Patricia Kraft ◽

Clifford Longley ◽

...

Keyword(s):

Therapeutic Efficacy ◽

Multiple Dose ◽

Hodgkin’S Lymphoma ◽

Xenograft Model ◽

Scid Mice ◽

Hodgkin's Lymphoma ◽

Non Hodgkin’S Lymphoma ◽

Non Hodgkin's Lymphoma ◽

Xenograft Models

Abstract Background: The clinical utility of SN38 (10-hydroxy-7-ethyl-camptothecin), the active moiety of CPT-11 (Camptosar®), has been severely compromised due to its poor solubility. EZN-2208 is a novel, water-soluble, polyethyleneglycol (PEG)-SN38 conjugate generated by linking SN38 with a multi-arm high molecular weight PEG, 40k 4-arm-PEG, via a glycine linker. Here, we compare the efficacy of EZN-2208 and CPT-11 in the treatment of animals bearing non-Hodgkin’s lymphomas. Methods: The in vitro cytotoxicity of EZN-2208 and CPT-11 were tested in human lymphoma (Raji, Daudi, DoHH2) cancer cell lines using the tetrazolium assay. The pharmacokinetics and maximum tolerated dose (MTD) of EZN-2208 and CPT-11 were evaluated in tumor-free severe combined immunodeficient (SCID) mice and the therapeutic efficacy was evaluated in xenograft models of non-Hodgkin’s lymphoma (Raji and Daudi). Results: In vitro, the IC50 of EZN-2208 ranged from 2–20nM and the in vitro potency of EZN-2208 was 30 to 50-fold more than CPT-11. The MTDs of EZN-2208 and CPT-11 in SCID mice were 30 and 60 mg/kg, respectively, when injected as a single dose. When administered in a multiple dose regimen (q2d x 5), the MTDs of EZN-2208 and CPT-11 were 10- and 40 mg/kg respectively. The pharmacokinetic profile of EZN-2208 was biphasic showing a rapid distribution phase (t1/2a =0.6h) and a slow elimination phase (t1/2b =19.3h for conjugate and 14.2h for SN38). In the Raji xenograft model, treatment with a single MTD of EZN-2208 resulted in a 500% improvement in life span (ILS) and 50% cures of animals compared with 19% ILS observed for mice treated with a single MTD of CPT-11. Multiple dose treatment of EZN-2208 resulted in 90% cures of animals compared with 63% ILS and no cures observed for the CPT-11 group. In the Daudi xenograft model, a single injection of EZN-2208 given as early treatment resulted in cures of 100% of animals in contrast to 66% ILS and no cures observed for CPT-11 group. When treatment was delayed, a single MTD of EZN-2208 caused 90% cures of animals, whereas CPT-11 treatment was completely ineffective. Conclusions: EZN-2208 demonstrated excellent efficacy in preclinical models of non-Hodgkin’s lymphoma. Ongoing Phase I studies will determine the optimal dose and schedule of EZN-2208.

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Incorporation of disinfectants for obtaining dental stone: microbiological and dimensional evaluation

Revista de Odontologia da UNESP ◽

10.1590/1807-2577.1036 ◽

2015 ◽

Vol 44 (1) ◽

pp. 24-30

Author(s):

Patrícia Lins Azevedo do Nascimento ◽

Rafael Bezerra Ribeiro ◽

Cícero Romão Gadê-Neto ◽

Alexandre Henrique de Moura Dias

Keyword(s):

Confidence Interval ◽

Metallic Matrix ◽

Dimensional Change ◽

Wilcoxon Test ◽

In Vivo Study ◽

In Vitro Test ◽

Type Iv ◽

Significant Difference

AIM: To assess dimensional change and antimicrobial activity of disinfectants substances incorporated during the dental stone manipulation. MATERIAL AND METHOD: In vivo - microorganisms were collected in alginate molds of 30 volunteers inoculated on BHI agar and incubated at 37 °C for 24 hours. The molds were cast with type IV gypsum, manipulated with saline (G1), 1% sodium hypochlorite (G2) and 4% chlorhexidine (G3), replacing the water. After setting of plaster with 1 hour two collections on models were made. After 24 hours, the readings were performed. The Kruskal-Wallis and Wilcoxon tests with confidence interval of 99% and 95% respectively were used. In vitro - Müeller Hinton agar petri dishes were inoculated with S. mutans (ATCC25175), S. sanguis (ATCC10556) and E. faecalis (ATCC29212), over which were placed steel rings filled with the same substances of the in vivo study. After deposition of gypsum and incubation, halos were measured with a digital caliper and data were submitted to ANOVA and Tukey's test with confidence interval of 95%. Dimensional Change - With a metallic matrix and a perfectly adapted tray, the insertion axis and force used for moulding and obtain 30 specimens in type IV gypsum were standardized, following the same distribution of the study groups in vivo. The specimens were measured by Image Pro Plus software and data were submitted to ANOVA and Tukey's test with confidence interval of 95%. RESULT: Data from the in vivo study demonstrated a significant difference between the mold and each model (p<0.001). In the Wilcoxon test there was no significant difference between groups of models. At the in vitro test, G2 showed greater inhibition zones in all micro-organisms tested compared to G3, but with respect to dimensional changes, there was a significant difference between solutions and metallic standard, where G3 caused less change than G2. CONCLUSION: Chlorhexidine 4% showed to be the most suitable disinfectant.

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Correlation of response of aplastic anemia patients to antilymphocyte globulin with in vitro lymphocyte stimulatory effect: predictive value of in vitro test for clinical response

Blood ◽

10.1182/blood.v77.10.2225.2225 ◽

1991 ◽

Vol 77 (10) ◽

pp. 2225-2230 ◽

Cited By ~ 8

Author(s):

T Abe ◽

H Matsuoka ◽

S Kojima ◽

Y Kamachi ◽

I Tsuge ◽

...

Keyword(s):

Aplastic Anemia ◽

Clinical Response ◽

Proliferative Response ◽

Mononuclear Cells ◽

In Vitro Test ◽

Antilymphocyte Globulin ◽

Peripheral Blood Mononuclear ◽

Color Flow ◽

Significant Difference

Abstract Therapy with antilymphocyte globulin (ALG) has been shown to be effective in restoring hematopoiesis to some patients with aplastic anemia. It would be useful to have a method for predicting those likely to be responders versus nonresponders. The mode of immunostimulatory action of ALG is of interest in addition to its immunosuppressive action. We examined in vitro the distribution of the proliferative responses of ALG-stimulated peripheral blood mononuclear cells (PBMCs) obtained from 18 patients with aplastic anemia, eight of whom responded to ALG and 10 who did not. We found a significant difference in the proliferative response of PBMCs obtained from the eight responders versus the 10 nonresponders (P less than .01). Two-color flow cytometry analysis of the patients' PBMCs stimulated by ALG in vitro showed that the CD4-positive subsets were activated to a greater extent by ALG than the CD8-positive subsets. Moreover, a positive correlation with the clinical response of patients to ALG with granulocyte-macrophage colony- stimulating factor produced by their PBMCs stimulated by ALG suggests that the immunostimulatory property of ALG has an important role in the treatment of aplastic anemia. Our results suggest that the clinical response to ALG therapy is correlated with its lymphocyte proliferative effect in vitro, and indicates that the assessment of the proliferative response of PBMCs in vitro would be useful in predicting the clinical response to ALG therapy.

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Correlation of response of aplastic anemia patients to antilymphocyte globulin with in vitro lymphocyte stimulatory effect: predictive value of in vitro test for clinical response

Blood ◽

10.1182/blood.v77.10.2225.bloodjournal77102225 ◽

1991 ◽

Vol 77 (10) ◽

pp. 2225-2230 ◽

Cited By ~ 1

Author(s):

T Abe ◽

H Matsuoka ◽

S Kojima ◽

Y Kamachi ◽

I Tsuge ◽

...

Keyword(s):

Aplastic Anemia ◽

Clinical Response ◽

Proliferative Response ◽

Mononuclear Cells ◽

In Vitro Test ◽

Antilymphocyte Globulin ◽

Peripheral Blood Mononuclear ◽

Color Flow ◽

Significant Difference

Therapy with antilymphocyte globulin (ALG) has been shown to be effective in restoring hematopoiesis to some patients with aplastic anemia. It would be useful to have a method for predicting those likely to be responders versus nonresponders. The mode of immunostimulatory action of ALG is of interest in addition to its immunosuppressive action. We examined in vitro the distribution of the proliferative responses of ALG-stimulated peripheral blood mononuclear cells (PBMCs) obtained from 18 patients with aplastic anemia, eight of whom responded to ALG and 10 who did not. We found a significant difference in the proliferative response of PBMCs obtained from the eight responders versus the 10 nonresponders (P less than .01). Two-color flow cytometry analysis of the patients' PBMCs stimulated by ALG in vitro showed that the CD4-positive subsets were activated to a greater extent by ALG than the CD8-positive subsets. Moreover, a positive correlation with the clinical response of patients to ALG with granulocyte-macrophage colony- stimulating factor produced by their PBMCs stimulated by ALG suggests that the immunostimulatory property of ALG has an important role in the treatment of aplastic anemia. Our results suggest that the clinical response to ALG therapy is correlated with its lymphocyte proliferative effect in vitro, and indicates that the assessment of the proliferative response of PBMCs in vitro would be useful in predicting the clinical response to ALG therapy.

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“Lemon grass oil mouth wash- A natural alternative”

Journal of Indian Dental Association ◽

10.33882/jida.13.25058 ◽

2019 ◽

Author(s):

Rohit Shah ◽

Rutuja Donde ◽

Dipika Mitra

Keyword(s):

Porphyromonas Gingivalis ◽

Antibacterial Properties ◽

Prevotella Intermedia ◽

In Vitro Test ◽

Bacterial Strains ◽

Lemon Grass ◽

Significant Difference ◽

Lemongrass Oil ◽

Mouth Wash

Aim and Objective: The objective of the study was to evaluate the anti-plaque efficacy of lemongrass oil mouthwash. It also assessed the antibacterial properties of lemon grass oil against Prevotella intermedia (Pi), Porphyromonas gingivalis (Pg), and Aggregatibacter actinomycetemcomitans (Aa) in vitro. Materials and Methods: Forty-five periodontally healthy subjects between the age group of 18 and22 years were enrolled in a 4-day plaque re-growth study. Subjects were divided into three groups, i.e., 15 in each group. Mouthwashes were labelled as A (0.25% lemongrass oil mouthwash), B (0.2% Chlorhexidine mouthwash) and C (placebo). Subjects were advised to refrain from any kind of mechanical oral hygiene procedure for four days. Plaque index (PI) was evaluated at baseline and on the fifth day. In vitro testing of lemon grass oil mouthwash was done against strains of Prevotella intermedia, Porphyromonas gingivalis, and Aggregatibactor actinomycetemcomitans, to evaluate the antimicrobial concentration of lemon grass oil mouthwash. Results: PI significantly increased from day 1 to day 5 (P < 0.05) in Groups A, B, and C. In inter-group comparison, there was a statistically significant difference between the three groups. However, chlorhexidine showed superior antiplaque activity. Lemon grass also demonstrated antiplaque activity, however, not as superior as chlorhexidine. In vitro test, lemon grass showed effective inhibition against all three bacterial strains Pi, Pg, and Aa at 0.01% concentration. Conclusion: Lemongrass oil mouthwash potently inhibits plaque formation and hence can be used as a natural herbal alternative. Key Words: lemongrass oil, antiplaque, antibacterial, chlorhexidine.

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EFEKTIVITAS EKSTRAK BUAH SAWO, BAWANG PUTIH, DAUN ANDONG, DAN BUAH PARE TERHADAP PERTUMBUHAN BAKTERI ESCHERICHIA COLI

JURNAL MEDIA KESEHATAN ◽

10.33088/jmk.v7i2.238 ◽

2018 ◽

Vol 7 (2) ◽

pp. 144-149

Author(s):

Susiwati Susiwati

Keyword(s):

Escherichia Coli ◽

Inhibition Zone ◽

Garlic Extract ◽

In Vitro Test ◽

E Coli ◽

Significant Difference ◽

Vitro Test ◽

Inhibition Zones ◽

Escherichia Coli Bacteria

This research aims to determine the inhibition of sapodilla fruit, garlic, andong leaves and pare fruit toward the growth of escherichia coli bacteria. Antimicroba test used paper disc diffusion was in-vitro test. Sapodilla fruit, garlic, andong leaves and pare fruit were extracted by using maceration process. The extracts were tested on the growth of E- coli bacteria. The highest inhibition zone (6,7mm) was found in andong leaves extract. The highest inhibition zone was 8.3 mm, whereas the inhibition of pare fruit did not not provide the inhibitory zone. It can be concluded that garlic extract, sapodolla extract and decoction of andong leaves have highly inhibitory in vitro. Based on stastistical analysis, there was Significant difference betwen the effectiveness of garlic extract with a decoction of andong leaves but the effectivess of garlic extract with sapodilla extract was not meaningful. Whereas pare fruit did not give any inhibition zones. From the result of this research, the society can be encouraged to consume andong leaves or sapodilla fruit to treat diarrhea. In addition, garlic spices and pare fruit also can be used to overcome diarrhea, which is caused by the bacterium E. coli.

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LncRNA HOXD-AS1 functions as a ceRNA to promote hepatocellular carcinoma metastasis in zebrafish xenograft models

10.21203/rs.3.rs-232919/v1 ◽

2021 ◽

Author(s):

Ying Zhang ◽

Xiaolu Wang ◽

Feng Qin ◽

Shaochang Jia

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Lines ◽

Noncoding Rna ◽

Xenograft Model ◽

Cerna Network ◽

Xenograft Models ◽

Proliferation And Invasion ◽

Hcc Cells

Abstract Background: A few studies have shown that long noncoding RNA (lncRNA) HOXD cluster antisense RNA 1 (HOXD-AS1) plays an important role in hepatocellular carcinoma (HCC) metastasis as a competing endogenous RNA (ceRNA), but there is little in vivo evidence. This study aims to explore the zebrafish HCC xenograft as an in vivo metastasis model to verify the ceRNA network of HOXD-AS1. Methods: The quantitative reverse transcription PCR (qRT-PCR) assay was used to assess the expression level of HOXD-AS1 in HCC cell lines. Knockdown of HOXD-AS1 or miR-130a-3p was performed by transfecting small interfering RNA (siRNA) or microRNA (miRNA) inhibitor, respectively. The proliferation and invasion of HCC cells in vitro were analyzed by CCK-8 and transwell assays. The growth and metastasis of HCC cells in vivo were assessed by zebrafish xenograft models.Results: We verified that HOXD-AS1 was overexpressed in all tested HCC cell lines than the normal hepatic cells. Silence of HOXD-AS1 suppressed cell proliferation and invasion in Hep3B and Huh7 HCC cell lines in vitro. In zebrafish xenograft models, knockdown of HOXD-AS1 also reduced the growth and metastasis of the two HCC cells. Moreover, downregulation of miR-130a-3p not only increased the HCC metastasis, but also rescued the metastasis which inhibited by silence of HOXD-AS1 in vitro and in vivo.Conclusions: Our study demonstrates the metastasis role of the HOXD-AS1/miR-130a-3p ceRNA network in HCC cells in vitro and in vivo, and these findings suggest that zebrafish xenograft model could be used for ceRNA mechanism verification in tumor metastasis.

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