This study investigated the relationship between women’s gender roles and their leadership in the context of Chinese higher education. Based on a questionnaire survey of 438 female leaders at various levels in Chinese higher education institutions, the research results show that gender-related factors such as family-role orientation and gender self-confidence have a close correlation with leadership aspiration and leadership self-efficacy. The results show that women’s family-role orientation has a negative effect on women’s leadership aspirations and leadership self-efficacy, while women’s gender self-confidence does not have a positive effect on leadership aspirations and leadership self-efficacy as hypothesized. Both family-role orientation and gender self-confidence have a close correlation with leadership aspirations. Leadership aspiration significantly mediated the effects between family-role orientation and leadership self-efficacy. These findings revealed the characteristics and changes in the relationship between gender and leadership among women leaders in Chinese higher education.