Anchorage-independent growth and the expression of cellular proto-oncogenes in normal human epidermal keratinocytes and in human squamous cell carcinoma cell lines.

The expression of multiple cellular proto-oncogenes and the in vitro anchorage-independent growth of normal human epidermal keratinocytes and several human squamous cell carcinoma cell lines were studied and correlated. Squamous cell carcinoma cell lines KB, Si Ha, HEp-2, and Fa Du showed high anchorage independency, and MS 751 and A-253 cell lines had minimum independency. However, the normal keratinocytes and the A-431 cell line did not show anchorage-independent growth. Both the normal human epidermal keratinocytes and cancer cell lines expressed multiple proto-oncogenes such as src, erb B-1, abl, fos, raf, H-ras, and myc, and the amount of expression of these oncogenes was notably higher in the cancer cell lines than in the normal keratinocytes. The expression of proto-oncogenes from the monolayer cultures of the cancer cell lines is poorly correlated with the anchorage independency of the cells. These data indicate that the anchorage independency is not directly linked to the expression of specific cellular proto-oncogene(s) of the monolayer cancer cell cultures.

Herpes simplex virus enhances the 7,12-dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis and amplification and overexpression of c-erb-B-1 proto-oncogene in hamster buccal pouch epithelium.

We studied the effects of herpes simplex virus type 1 (HSV-1) inoculation and topical 7,12-dimethylbenz[a]anthracene (DMBA) application, alone or in combination, on the carcinogenesis and on the amplification and expression of various cellular proto-oncogenes in hamster buccal pouch tissue. Topical DMBA treatment produced tumor formation in pouches, but HSV-1 inoculation, alone caused no neoplastic changes. In pouch tissues receiving both DMBA application and HSV-1 inoculation, the development of initial leukoplakia and tumor has hastened and enhanced in comparison with those receiving DMBA alone. Topical DMBA application to pouch tissue induced an amplification and an increase in the expression of cellular erb-B-1 (c-erb-B-1) proto-oncogene in the epithelial tissue, whereas repeated infection with HSV-1 alone did not. Topical DMBA combined with HSV-1 inoculation, however, resulted in greater amplification and expression of c-erb-B-1 proto-oncogene in the pouch epithelial tissue compared to the DMBA alone. These data indicate that HSV-1 inoculation significantly increases the carcinogenic activity of DMBA, in part, by probably enhancing DMBA-induced amplification and expression of c-erb-B-1 proto-oncogene in hamster buccal pouch tissue.

Primary structure of the polymerase acidic (PA) gene of an influenza B virus (B/Sing/222/79).

A DNA copy of influenza B/Singapore/222/79 viral RNA segment 3 containing the gene coding for the polymerase acidic (PA) protein has been cloned in Escherichia coli plasmid pBR322, and its nucleotide sequence has been determined. The cDNA clone was incomplete and contained 1810 nucleotides (nt 396 to 2205). The remaining nucleotide sequence at both 5' and 3' ends of B PA gene was obtained by sequencing the viral RNA (minus sense) and messenger RNA (plus sense) using oligonucleotide primers. The influenza B PA gene contains 2304 nucleotides and codes for a protein of 725 amino acids with a molecular weight of 83,000. The predicted influenza B PA protein is less acidic than the influenza A PA protein. Computer alignment of the influenza B PA amino acid sequence with that of influenza A PA (A/PR/8/34) revealed an overall 38% direct homology which increases to 45% in the carboxyl terminus half of the protein. In addition, comparison of the secondary structural elements, hydropathy profile, and isofunctional amino acid changes between B PA and A PA proteins demonstrated a strong structural and possibly functional conservation between these two proteins. These data suggest that PA genes of influenza A and B viruses arose from a common ancestor gene.