Use of the membrane invasion culture system (mics) as a screen for anti‐invasive agents

Abstract

The Membrane Invasion Culture System (MICS) assay was adapted for relatively rapid screening of compounds and used to identify anti-invasive drugs that inhibit human and murine tumor cell migration through a reconstituted basement membrane in vitro. Cell lines demonstrating low and high invasive and metastatic potentials were tested with all compounds for tumoricidal effects prior to evaluation in MICS at non-cytotoxic doses. The effect on invasive potential in the MICS assay was determined in 3 categories: (1) 48 hr drug pre-treatment prior to seeding in the MICS (exceptions: 90 min pre-treatment with pertussis toxin and, for some studies, continuous exposure for 2-7 days); (2) peptide or prostaglandins 2 hr after seeding and attachment to the membranes in MICS followed by continuous exposure; and (3) cells receiving neither drug nor peptide treatment and serving as controls in each MICS chamber. Since invasion involves cellular motility and deformability, some cytoskeleton disrupting agents were selected. Of these, vincristine, colcemid and colchicine inhibited invasion but taxol did not. Pre-treatment with cAMP agonists produced conflicting results: dibutyryl cAMP and 8-(4-chloro-phenylthio) cAMP resulted in 50% and 38% reduction in invasion, respectively, whereas 8-bromo cAMP stimulated invasive potential by 30%. Forskolin and cholera toxin both significantly reduced invasiveness. Pre-treatment with 5-azacytidine and araC, to consider the role of methylation and proliferations decreased invasive ability. Anti-metastatic drugs such as gamma-interferon and razoxane inhibited invasive potential but to varying degrees. Treatment of cells with prostaglandins E2, F2 alpha, A2, and D2 were ineffectual; however, indomethacin mildly inhibits invasion (less than 30%).(ABSTRACT TRUNCATED AT 250 WORDS)

Transforming growth factor beta stimulates mammary adenocarcinoma cell invasion and metastatic potential.

“…Total number of invading cells was calculated based upon the surface area per high-power field and the surface area of the Minifold-well. Percent invasion was then calculated as described (32 added concomitantly with TGF-,8 to MTLn3 cells, the number of lung colonies was similar to controls, indicating that the metastasis-enhancing factor was, indeed, TGF-P (Fig. 1).…”

“…To measure tumor cell invasion, the membrane invasion culture system (MICS) (32,33) was used with minor modifications. Instead of the 12/15 well configuration used in the original MICS, we used a 24-well manifold chamber fitted with a silicon gasket between the upper and lower plates to ensure no lateral crosscontamination between wells (Neuroprobe, Cabin John, MD).…”

Extracellular Matrix Invasion in Metastases and Angiogenesis: Commentary on the Matrigel “Chemoinvasion Assay”

“…This model was optimized to fit the needs of individual laboratories and to run several samples at the same time, and it has been adapted to commercial packages. Screening systems, first employed by Mary Hendrix as the MICS assay (8), are also commercially available.…”

A new superinvasive in vitro phenotype induced by selection of human breast carcinoma cells with the chemotherapeutic drugs paclitaxel and doxorubicin

“…While a few reports indicate that paclitaxel exposure increases motility and invasion (Welch et al, 1989;Silbergeld et al, 1995), others indicate an inhibitory effect on invasion (Westerlund et al, 1997;Sgadari et al, 2000). Silbergeld et al (1995) reported that increasing the paclitaxel concentration caused increased locomotion in glioblastoma cell lines.…”

“…Silbergeld et al (1995) reported that increasing the paclitaxel concentration caused increased locomotion in glioblastoma cell lines. Welch et al (1989) showed that pretreatment of cell lines, with low and high invasive potentials, with vincristine, colcemid, or colchicine (but not paclitaxel), at noncytotoxic levels, resulted in inhibition of invasion. Westerlund et al (1997) showed that OVCAR-3 cell attachment, migration and in vitro invasion were significantly decreased after paclitaxel treatment.…”