[Preliminary application of mRNA differential display technique in the isolation of genes possibly associated with tumor metastasis].
We have previously reported that in vitro treatment with interferons of MA891, a spontaneous mouse mammary tumor capable of metastasizing to the lungs upon subcutaneous inoculation, could modulate its metastatic potential. IFN-gamma increases while IFN-alpha decreases lung metastasis. Identifying the genes differentially expressed in MA891 cells separately treated with IFN-alpha and IFN-gamma. MA891 cells were treated in vitro with 200 U/ml IFN-alpha or IFN-gamma respectively for 48 h. Total RNA was isolated with the single step method of acid guanidinium thiocyanate. After being digested by DNase I, the two RNA samples were differentially displayed with T12MA/T/C/G x 20 arbitrary primer pairs. Most of the RT-PCR were done twice. The differentially expressed cDNA bands were isolated, reampliated and cloned. Ten differentially expressed cDNA bands were identified: T2 alpha, T2 gamma, C4 alpha, T6 alpha, T11 gamma, G15 gamma, T16 gamma, T5 alpha 1, T5 alpha 2, T17 alpha, T15 alpha, T15 gamma, named according to the type of IFN and primer pairs used. The total RNA samples of MA891 cells treated by IFN-alpha and IFN-gamma were differentially displayed with T12MA/T/C/G x 20 arbitrary primer pairs. Theoretically they can cover 95% mRNAs. It is anticipated that one or more of the ten cDNA clones may be related to the metastatic potential of MA891 cells.
Zhang
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Q
Cancer Institute, CAMS, Beijing.
Zhang
Y
Y
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics; D007370:Interferon Type I / Q000494:pharmacology*; D007371:Interferon-gamma / Q000494:pharmacology*; D008175:Lung Neoplasms / Q000556:secondary; D008325:Mammary Neoplasms, Experimental / Q000235:genetics* / Q000473:pathology; D051379:Mice; D009362:Neoplasm Metastasis / Q000235:genetics; D009857:Oncogenes; D016133:Polymerase Chain Reaction / Q000379:methods; D012334:RNA, Neoplasm / Q000235:genetics; D011994:Recombinant Proteins; D014407:Tumor Cells, Cultured / Q000187:drug effects
D018076:DNA, Complementary; D007370:Interferon Type I; D012334:RNA, Neoplasm; D011994:Recombinant Proteins; D007371:Interferon-gamma