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Estimation of interleukin 6 production by reverse transcriptase-polymerase chain reaction in four human myeloma cell
A Suzuki1, T Takahashi, Y Okuno
1Second Department of Internal Medicine, Kyoto University, Japan.
Abstract:
Although an autocrine growth mechanism through interleukin 6 has been advocated in human myeloma cells, reports of IL-6 production by cells from established myeloma cell lines are rare. In the present study, we examined whether or not a minute amount of interleukin 6 is produced in 4 human myeloma cell lines. IL-6 production was not detected in any of the 4 lines by enzyme immunoassay, bioassay with two interleukin 6-dependent murine hybridoma cell lines and Northern hybridization. However, we detected interleukin 6 mRNA in one (U266) of the 4 lines by the reverse transcriptase-polymerase chain reaction. Nevertheless, the proliferation of all 4 lines was not inhibited by an anti-interleukin 6 antibody. These results suggest that autocrine stimulation by interleukin 6 is not involved in the majority of human myeloma cell lines.
Insights
Interleukin 6 (IL-6) autocrine growth is unlikely in most human myeloma cell lines. Researchers found minimal IL-6 mRNA in one cell line but no detectable IL-6 protein or growth inhibition by antibodies.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Interleukin 6 (IL-6) is implicated in autocrine growth mechanisms in human myeloma cells.
- However, evidence of IL-6 production by established myeloma cell lines is scarce.
Purpose of the Study:
- To investigate the production of IL-6 in four human myeloma cell lines.
- To determine if IL-6 autocrine stimulation contributes to myeloma cell proliferation.
Main Methods:
- Enzyme immunoassay, bioassay using IL-6-dependent hybridoma cell lines, and Northern hybridization were employed to detect IL-6.
- Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect IL-6 mRNA.
- The effect of an anti-IL-6 antibody on cell proliferation was assessed.
Main Results:
- IL-6 production was undetectable in all four myeloma cell lines by immunoassay, bioassay, and Northern hybridization.
- IL-6 mRNA was detected in one cell line (U266) via RT-PCR.
- Anti-IL-6 antibody did not inhibit the proliferation of any of the four cell lines.
Conclusions:
- Autocrine stimulation by IL-6 does not appear to be a significant factor in the growth of most human myeloma cell lines.
- While IL-6 mRNA can be present, functional IL-6 protein production or its role in proliferation may be limited.