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Laminin Suppresses Progesterone Production by Human Luteinizing Granulosa Cells via Interaction with Integrin ₆ß₁¹

Department of Gynecology and Obstetrics, Faculty of Medicine (H.F., T.H., K.N., S.Y., T.M.), Institute for Virus Research (M.U.), and Chest Disease Research Institute (M.M.), Kyoto University, Sakyo-ku, Kyoto 606–01, Japan

Address all correspondence and requests for reprints to: Hiroshi Fujiwara, M.D., Department of Gynecology and Obstetrics, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto, 606–01, Japan.

We previously raised a murine monoclonal antibody (mAb), OG-1, against human granulosa cells (GC) and reported that human GC express the OG-1 antigen with the highest immunoreactivity during the periovulatory phase. Later, we showed that the OG-1 antigen is identical to human integrin ₆, and that human GC express integrin ₆ß₁, but not ₆ß₄.

In the present study, we examined the expression of laminin (LN), the ligand for integrin ₆ß₁. Flow cytometry showed that LN was bound to the cell surface of some GC obtained from preovulatory follicles of patients undergoing in vitro fertilization. Immunohistochemistry showed that LN was detected between luteinizing GC in the early corpora lutea.

To examine the effect of LN on steroidogenesis by human luteinizing GC, GC obtained from patients undergoing in vitro fertilization were cultured on mouse LN-coated or noncoated plastic dishes in medium containing 5% FCS for 24 h. In the absence or presence of hCG (1 IU/mL), GC cultured on LN-coated dishes produced 0.70- and 0.67-fold less progesterone than those on noncoated dishes, respectively (P < 0.05).

We examined the effect of the interaction of integrin ₆ß₁ and LN on steroidogenesis by human luteinizing GC. We cultured GC with 5 µg/mL of the anti-₆ mAb GoH3, which inhibits the interaction between human integrin ₆ß₁ and mouse LN, or with a control rat mAb (TER199) on mouse LN-coated dishes in serum-free medium for 24 h. In the absence or presence of hCG (1 IU/mL), GC cultured with GoH3 produced 1.97- and 1.94-fold more progesterone than the control cells (P < 0.01 and P < 0.05, respectively). In contrast, when GC were cultured on dishes coated with type IV collagen, progesterone production was not enhanced by GoH3. Furthermore, the anti-₆ mAb OG-1, which does not inhibit the interaction between integrin ₆ß₁ and LN, had no effect on the progesterone production by GC cultured on LN.

These results indicate that LN suppresses the luteinization of human luteinizing GC via integrin ₆ß₁ and that integrin ₆ß₁ regulates the luteinization of human GC during the periovulatory phase.

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