Substitution of Chloroform by Bromochloropropane in the Single-Step Method of RNA Isolation

Chomczyński, Piotr; Mackey, Karol

doi:10.1006/abio.1995.1126

Cited by 187 publications

(110 citation statements)

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“…The starting material for reverse transcription-PCR (RT-PCR) was total RNA, which was isolated from PBS-washed cells that were solubilized with a guanidine isothiocyanate-based buffer (Tri-Reagent, SigmaAldrich, St. Louis, MO), according to a modification of the method of Chomczynski (18). Total RNA (5 g) was used as the template for cDNA synthesis, using the First Strand kit of Novagen (Madison, WI) according to the manufacturer's instructions, with 5% of the resulting cDNA product being used for each PCR reaction.…”

Section: Methodsmentioning

confidence: 99%

CD74 Is Expressed by Multiple Myeloma and Is a Promising Target for Therapy

Burton

Ely

Reddy

et al. 2004

Clinical Cancer Research

142

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Purpose: CD74 (HLA-DR-associated invariant chain) plays a role in antigen presentation. In addition to its expression on antigen-presenting cells, it is expressed by carcinomas of renal, lung, gastric, and thymic origin and by certain sarcomas. The restricted expression of CD74 by normal tissues and its very rapid internalization make CD74 an attractive therapeutic target for both cancer and immunologic diseases. Preclinical efficacy of anti-CD74 monoclonal antibody (mAb) therapy has been demonstrated in B-lymphoma models. Because there are few validated antigenic targets in multiple myeloma, CD74 expression was examined.Experimental Design: CD74 expression was assessed by immunohistochemistry in bone marrow biopsies of known multiple myeloma cases. Its expression was measured by flow cytometry in multiple myeloma lines, and CD74 mRNA expression was determined by reverse transcription-PCR. In addition, the in vitro antiproliferative effect of LL1 mAb was evaluated on a CD74؉ multiple myeloma cell line using a [ 3 H]thymidine incorporation assay.Results: CD74 expression was observed in 19 of 22 cases of multiple myeloma, with most expressing moderate to high levels in the majority of malignant plasma cells. CD74 was expressed by most multiple myeloma cell lines, as was CD74 mRNA, at levels mirroring CD74 protein. Also, unlabeled LL1 mAb mediated in vitro growth inhibition of a CD74؉ multiple myeloma cell line.Conclusions: CD74 expression is frequent in multiple myeloma, with predominant expression by the malignant plasma cells. Because anti-CD74 mAbs internalize very rapidly and LL1 mAb has shown efficacy in B-lymphoma models, CD74 represents a novel and promising target for treatment of multiple myeloma. Therefore, LL1 mAb is well suited as a carrier of radionuclides, drugs, or toxins, and also has activity as an unlabeled mAb, thereby supporting its development for this unmet need in cancer therapy.

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Section: Methodsmentioning

confidence: 99%

CD74 Is Expressed by Multiple Myeloma and Is a Promising Target for Therapy

Burton

Ely

Reddy

et al. 2004

Clinical Cancer Research

142

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“…Total RNA was isolated from microdissected and homogenized maxilloturbinates and pulmonary axial airways by following the method of Chomczynski and co-workers (55,56). Steady-state levels of rat mucin 5AC (rMuc5AC) mRNA were then determined in RNA samples using a quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique.…”

Section: Protocols and Methodologic Approaches For Assessing Mucous Cmentioning

confidence: 99%

Effects of Ozone and Endotoxin Coexposure on Rat Airway Epithelium: Potentiation of Toxicant-Induced Alterations

Wagner¹,

Hotchkiss²,

Harkema³

2001

Environmental Health Perspectives

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“…Total RNA was extracted from the cultured cells with the acid guanidinium thiocyanate-phenolchloroform extraction method (Chomczynski and Mackey, 1995). Northern blot analysis was performed according to a previously described method (Choi et al, 2003).…”

Section: Northern Blot Analysismentioning

confidence: 99%

Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death

Choi

Yoon²,

Park³

et al. 2007

Exp Mol Med

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Although N-myc downstream regulated gene 2 (NDRG2) has been known to be a tumor suppressor gene, the function of this gene has not been elucidated. In the present study, we investigated the expression and function of NDRG2 in human gastric cancer. Among seven gastric cancer and two non-cancer cell lines, only two gastric cancer cell lines, SNU-16 and SNU-620, expressed NDRG2, which was detected in the cytoplasm. Interestingly, NDRG2 was highly expressed in normal gastric tissues, but gastric cancer patients were divided into NDRG2-positive and -negative groups. The survival rate of NDRG2-negative patients was lower than that of NDRG2-positive patients. We confirmed that the loss of NDRG2 expression was a significant and independent prognostic indicator in gastric carcinomas by multivariate analysis. To investigate the role of NDRG2 in gastric cancer cells, we generated a NDRG2-silenced gastric cancer cell line, which stably expresses NDRG2 siRNA. NDRG2-silenced SNU-620 cells exhibited slightly increased proliferation and cisplatin resistance. In addition, inhibition of NDRG2 decreased Fas expression and Fas-mediated cell death. Taken together, these data suggest that inactivation of NDRG2 may elicit resistance against anticancer drug and Fas-mediated cell death. Furthermore, case studies of gastric cancer patients indicate that NDRG2 expression may be involved in tumor progression and overall survival of the patients.

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Substitution of Chloroform by Bromochloropropane in the Single-Step Method of RNA Isolation

Cited by 187 publications

References 0 publications

CD74 Is Expressed by Multiple Myeloma and Is a Promising Target for Therapy

CD74 Is Expressed by Multiple Myeloma and Is a Promising Target for Therapy

Effects of Ozone and Endotoxin Coexposure on Rat Airway Epithelium: Potentiation of Toxicant-Induced Alterations

Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death

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