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Table 1 Summary of Anti-Mammastatin Immunoblot and Inhibition Assay

From: Differential Expression of Mammastatin in Normal and Breast Cancer Cells

   Species Expression Inhibition of Growth
Cell Type N 44 kD 49/53 kD Yes/No (#)
Normal, primary cultures 133 +/- ++++ Y(2/2)
Normal breast tissue* 5 + ++ NT
Mammary cell line 16 + (11/16) - Y (12/12)
Breast tumor lysate 25 + (17/25) - NT
Non-Mammary cell lines 8 + (3/8) ** - N (0/8)
  1. Analysis of Mammastatin expression by immunoblot: Lysatesof fresh tissue, primary cultures and cell lines were analyzed by 7G6.6,anti-mammastatin, immunoblot. N = number of individual samples repeated at least3 times. (++++) intense expression, (-) no expression, NT = not treated.
  2. Inhibition Assays: Cells were plated in growth media at 104cells/ml and treated with 10 ng/ml purified Mammastatin or control. Culturescounted by Coulter Counter after six days culture.
  3. Carcinoma cells in which mammastatin was detected: MCF-7,ZR-75-1, T47D, Sk Br 3, ZR-75-30, BT 475, UACC 893, MDA-MB-361, Sum 44, Sum 52,Sum 102 (Sum lines from S. Either, Uof Mich), Hep G2, OvCar, BxPC3, andA253.
  4. Carcinoma cells in which mammastatin was not detected:MDA-MB-231, BT-20, MDA-MB-435, HBL-100, Du 4475, UM 17B (squamous line from T.Corbett Uof Mich), SK Mel 31, 293T, HT 1080. * Two additional normaltissue lysates from breast cancer patients showed no Mammastatin expression.** Similar size proteins (between 40 and 55 kD) were identified inBxPc3, HepG2 and OvCar, A253 cells but these cell lines did not producedinhibitory activity.