Chemosensitivity testing of fresh human gastric cancer with highly purified tumour cells using the MTT assay

Yamaue, Hiroki; Tanimura, Hiroshi; Noguchi, Kohei; Hotta, Tsukasa; Tani, Masaji; Tsunoda, Takuya; Itō, Makoto; Tamai, Mikiko; Iwakura, Shinji

doi:10.1038/bjc.1992.362

Cited by 69 publications

(57 citation statements)

References 19 publications

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“…Methods using mechanical or enzymatic dissociation of tumoral fragments to isolate and then cultivate tumor cells suspensions are time consuming, expensive, and success rate do not exceed 50%. 32,33 Using poly-HEMA-coated surface we have established a simple and cost-effective model were tumor fragments can be cultured for up to 4 days while keeping their three-dimensional architecture and their proliferative activity, without bacterial contamination or fibroblastic proliferation. 34 The use of a double immunostaining to exclude nontumorous proliferative cells and a computerized count of proliferative tumor cells allowed an easy and reproducible comparison of proliferation in cultured fragments.…”

Section: Discussionmentioning

confidence: 99%

Evidence of heterogeneity within colorectal liver metastases for allelic losses, mRNA level expression and in vitro response to chemotherapeutic agents

Goasguen

Chaisemartin

Brouquet

et al. 2010

Intl Journal of Cancer

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A goal of oncology is to predict chemosensitivity of tumors. This approach assumes that in a patient all tumor deposits are homogeneous. We have tested the heterogeneity between several samples of the same liver metastasis (LM; intrametastatic heterogeneity) or between multiple LM (intermetastatic heterogeneity) from colorectal cancer in a single patient. In 16 untreated patients, several fragments of LM and nontumorous liver were collected. Heterogeneity to anticancer drug treatment was assessed in vitro on primary tissue cultures on poly-HEMA-coated surface with or without the topoisomerase-I inhibitor metabolite SN-38. Heterogeneity of response to SN-38 was observed in 55% of cases from one fragment to another in the same LM and in 64% of cases from one LM to another in the same patient. Allelic losses were characterized on 5q, 8p, 17p, 18q, 22q using 29 microsatellites markers. Seven patients (58%) had a perfect homogeneity for allelic losses in their LM whereas 3 (21%) had intrametastatic and 2 (18%) had intermetastatic heterogeneity. The analysis of gene expression was carried out by real time RT-PCR quantification using specific probes for TS, TOPO1, ERCC1, and CES2. Level expression of genes tested appeared heterogeneous with average variations of 57(623)%, 52(618)%, 53(618)%, 56(616)% for TS, TOPO1, ERCC1, and CES2 respectively for intermetastatic variability and 47(626)%, 36(614)%, 38(619)%, and 56(629)%, respectively for intrametastatic variability. Our results demonstrate intermetastatic and intrametastatic heterogeneity suggesting that pretherapeutic analysis of a single tumor biopsy is likely to lead to a misinterpretation of sensitivity to anticancer treatment.Colorectal carcinoma is one of the most prevalent cancer in western countries.1,2 Nearly 50% of patients suffering from colorectal cancer develop distant metastases at some point during the course of their disease, which represents the main cause of cancer related death. In case of liver metastases, treatment is based upon surgical resection but only 10-20% of metastases are amenable to resection.3 Patients with multiple liver metastases receive systemic chemotherapy.The increasing number of active cytotoxic drugs and targeted therapies has placed clinicians in front of a broad spectrum of therapeutic options. [4][5][6][7][8] Predictive factors for the response to chemotherapy are currently extensively studied in order to identify markers of chemosensitivity or chemoresistance which could guide oncologists for the choice of the most effective drug for a given patient. [9][10][11][12][13][14][15] To date, most of these studies aiming to determine a molecular signature of the tumor are conducted from DNA or RNA extracted from a single tumor biopsy either on primary tumor or metastases.Validity of such an approach predicting chemosensitivity is based on the clonality of molecular alterations within the tumor and metastases, in other word considering that all part of the tumor and all tumor deposits (i.e., primary and metastases) have the same m...

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Section: Discussionmentioning

confidence: 99%

Evidence of heterogeneity within colorectal liver metastases for allelic losses, mRNA level expression and in vitro response to chemotherapeutic agents

Goasguen

Chaisemartin

Brouquet

et al. 2010

Intl Journal of Cancer

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“…Following cellular adhesion, freshly prepared anticancer drugs, including VCR, 5-FU, CDDP, were added with the final concentration being 0.01-, 0.1-, 1-and 10-times the human peak plasma concentration for each drug, as previously described (19). The peak serum concentrations of various anticancer drugs are 0.5 µg/ml for VCR, 10 µg/ml for 5-FU and 2.0 µg/ml for CDDP (20,21).…”

Section: Introductionmentioning

confidence: 99%

Paeoniflorin modulates multidrug resistance of a human gastric cancer cell line via the inhibition of NF-κB activation

Liu

2011

Mol Med Report

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Abstract.Research into the evasion of drug resistance and adverse effects is highly significant and urgent in the clinic. Therefore, accumulating studies have focused on the development of novel target-specific molecules related to drug resistance, and the establishment of rational therapeutic approaches. Currently, studies have shown that NF-κB activation may play an essential role in the development of chemotherapy resistance in carcinoma cells. Paeoniflorin, a principal bioactive component of the root of Paeonia lactiflora Pall., has been reported to exhibit various pharmacological effects. In the present study, we reported for the first time that paeoniflorin at non-toxic concentrations may effectively modulate multidrug resistance (MDR) of the human gastric cancer cell line SGC7901/vincristine (VCR) via the inhibition of NF-κB activation and, at least partly, by subsequently downregulating its target genes MDR1,

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“…Fresh excised tumour tissues were processed by enzymatic digestion as described previously (Tani et al, 1995;Yamaue et al, 1991;Yamaue et al, 1992;Iwahashi et al, 1992). Briefly, tumour tissues were dissected into pieces smaller than 2 mm3 and these were immersed in medium containing collagenase (2 mg ml-', type V-S; Sigma, St Louis, MO, USA), hyaluronidase (10 U ml-', type IV-S; Sigma) and DNAase-I (0.4 mg ml-'; Sigma).…”

Section: Materials and Methods Patientsmentioning

confidence: 99%

Fibronectin promotes the proliferation of cytotoxic T lymphocytes generated from cancer patients

Mizobata

Tanimura²,

Yamaue³

et al. 1996

Br J Cancer

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Chemosensitivity testing of fresh human gastric cancer with highly purified tumour cells using the MTT assay

Cited by 69 publications

References 19 publications

Evidence of heterogeneity within colorectal liver metastases for allelic losses, mRNA level expression and in vitro response to chemotherapeutic agents

Evidence of heterogeneity within colorectal liver metastases for allelic losses, mRNA level expression and in vitro response to chemotherapeutic agents

Paeoniflorin modulates multidrug resistance of a human gastric cancer cell line via the inhibition of NF-κB activation

Fibronectin promotes the proliferation of cytotoxic T lymphocytes generated from cancer patients

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