Although the impact of cigarette smoking on glucose homeostasis has been extensively studied, the results, however, are still not conclusive. We, therefore, conducted a cross-sectional analysis of a non-diabetic Chinese cohort collected by the China Health and Nutrition Survey (CHNS 2009) to comprehensively assess the relationship between smoking, Hemoglobin A1c, beta-cell function and insulin sensitivity. The cohort included a total of 5965 individuals (47.4% male) with a mean age of 49.23 years, and 4140 of which were non-smokers (69.4%), 834 were current light smokers (13.9%) and 991 were current heavy smokers (16.6%). Current smokers were predominantly males (93.6%) with a lower BMI (22.95 versus 23.42 kg/m2). HbA1c levels were dose-dependently increased with smoking exposure (5.39%, 5.42% and 5.45%, respectively, P = 0.007). Non-smokers were served as a referent, the adjusted ORs for type 2 diabetes were 1.12 (P = 0.256, light smokers) and 1.26 (P = 0.014, heavy smokers), indicating a positive relationship between cigarette smoking and incidence of diabetes. HOMA%B was decreased in a dose-responsive manner with cigarette smoking (4.80, 4.79 and 4.76, P = 0.036), suggesting an adverse effect of smoking on beta-cell function. Collectively, cigarette smoking is dose-dependently associated with decreased HOMA%B, and current smokers were clearly in a higher risk for diabetes as manifested by the elevated HbA1c.