Sulphorhodamine B assay for measuring proliferation of a pigmented melanocyte cell line and its application to the evaluation of crude drugs used in the treatment of vitiligo

Lin, Zhi Xiu; Hoult, J.R.S.; Raman, Amala

doi:10.1016/s0378-8741(98)00199-8

Cited by 60 publications

(43 citation statements)

References 22 publications

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“…At these low concentrations, the solvents exhibited neither stimulatory nor inhibitory effects on the growth of these cell lines (data not shown). Cells were incubated for 72 h before their proliferation rates were determined by Sulphorhodamine B (SRB) assay using a method described elsewhere (Skehan et al, 1990;Lin et al, 1999). Briefly, at the end of the incubation, the cells were fixed by addition of 25 μL of cold 50% trichloroacetic acid (TCA, 4°C) on top of the 150 μL of growth medium already present in each well.…”

Section: Methodsmentioning

confidence: 99%

Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines

Lau

Lin

Zhao

et al. 2008

Phytotherapy Research

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Brucea javanica fruit is thought to have anticancer properties in Chinese medicine and its extract has been shown to possess antiproliferative and pro-apoptotic activities on human carcinoma cells. In the present study we demonstrated for the first time that Fructus Bruceae extract exhibited cytotoxic effects on the three pancreatic adenocarcinoma cell lines, PANC-1, SW1990 and CAPAN-1; the effects were comparable to those exhibited by camptothecin in our culture system. In addition, Fructus Bruceae extract induced fragmentation of genomic DNA, as evidenced by Hoechst staining and the cell death detection ELISA(PLUS) assay. Western blot analysis further showed down-regulation of pro-caspase 3 protein expression, indicating that the observed cytotoxic effects of the extract were associated with induction of apoptosis. These findings are not only significant in the development of traditional Chinese medicine as an alternative treatment for pancreatic cancer, but also in the elucidation of the potential mechanism(s) of Fructus Bruceae extract in cancer therapy.

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Section: Methodsmentioning

confidence: 99%

Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines

Lau

Lin

Zhao

et al. 2008

Phytotherapy Research

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“…K562/ADR and K562/S cells were cultured in PRMI1640 (Gibco) medium, supplemented with 10 fetal bovine serums at 37°C in a humidified incubator with 5% CO 2 . SRB assay was performed as described in literatures 15,16) : Cells were seeded into 96-well plates at 4000 viable cells per well. Blank wells and control wells were set up.…”

Section: Methodsmentioning

confidence: 99%

Cytotoxic Polyprenylated Xanthones from the Resin of Garcinia hanburyi

Han

Wang

Yang

et al. 2006

CHEMICAL & PHARMACEUTICAL BULLETIN

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“…The proliferation rates of Capan-2, WRL68 and PPC were determined by the SRB assay, a colorimetric end-point microplate assay which quantifies viable cells by staining their cellular protein content (10). BD was reconstituted in DMSO to produce a stock solution and then diluted with culture medium to various concentrations for cell culture experiments.…”

Section: Involvement Of the Mitochondrial Pathway In Bruceine Dmentioning

confidence: 99%

Involvement of the mitochondrial pathway in bruceine D-induced apoptosis in Capan-2 human pancreatic adenocarcinoma cells

et al. 2012

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Abstract. The fruit of Brucea javanica L. is a common herb used in Chinese medicine for the treatment of a variety of cancers. Our research group has previously identified bruceine D (BD), a quassinoid found abundantly in B. javanica, to have potent cytotoxic effect on a number of pancreatic cancer cell lines, including Panc-1, SW1990 and Capan-1 cells. In the present study, we showed that BD was also able to inhibit the growth of the Capan-2 human pancreatic adenocarcinoma cell line, but it exerted only modest cytotoxicity on the WRL68 human hepatocyte cell line and a human pancreatic progenitor cell line. The antiproliferative effects of BD were comparable to those exhibited by camptothecin and gemcitabine in our culture system. We found a dose-dependent decrease of the mitochondrial membrane potential in BD-treated Capan-2 cells as measured by the JC-1 assay. BD exposure was able to attenuate the expression of Bcl-2 protein in Capan-2 cells as detected by western blot analysis. In addition, the expression of both caspase 9 and caspase 3 in BD-treated Capan-2 cells was significantly accentuated. Moreover, BD was capable of inducing the fragmentation of genomic DNA in Capan-2 cells as evidenced by Hoechst staining. Cell cycle analysis demonstrated that BD could increase the percentage of Capan-2 cells in the subG 1 phase in a dose-related manner. An increase in the apoptosis of Capan-2 cells was also observed by Annexin V and PI staining. These results unequivocally indicate that BD induces cytotoxicity in Capan-2 cells via the induction of cellular apoptosis involving the mitochondrial pathway. IntroductionHuman pancreatic cancer is a gastrointestinal malignancy with extremely poor prognosis owing to a lack of early clinical diagnosis and its intrinsic resistance to radiotherapy and chemotherapy (1,2). According to the Hong Kong Cancer Registry in 2009 pancreatic cancer was the 8th and 6th major cause of cancer-related deaths in males and females, respectively. As for the management of pancreatic cancer, surgical resection remains the only potentially curative treatment of pancreatic cancer. However, only 5-25% of the patients diagnosed with pancreatic cancer were operable, and chemotherapy thus remains the mainstay of palliative management for the locally advanced and metastatic stage of this cancer. The median disease-free survival following complete resection of the pancreatic tumor and adjuvant chemotherapy with the first line chemotherapeutic agent gemcitabine is 13.4 and 6.9 months, respectively (3-6). Although camptothecin has a positive effect on arresting pancreatic tumor growth, many unwanted sideeffects limit its clinical application (5,7). All these dismal data highlight an urgent need for new therapeutic agents that could produce better clinical outcomes for this deadly disease.Our previous studies have shown that the fruit of Brucea javanica L., a Chinese medicinal plant commonly used for cancer treatment, has potent in vitro anti-pancreatic cancer activity (8). Furthermore, bruceine D (BD), a qu...

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Sulphorhodamine B assay for measuring proliferation of a pigmented melanocyte cell line and its application to the evaluation of crude drugs used in the treatment of vitiligo

Cited by 60 publications

References 22 publications

Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines

Brucea javanica fruit induces cytotoxicity and apoptosis in pancreatic adenocarcinoma cell lines

Cytotoxic Polyprenylated Xanthones from the Resin of Garcinia hanburyi

Involvement of the mitochondrial pathway in bruceine D-induced apoptosis in Capan-2 human pancreatic adenocarcinoma cells

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