Abstract Background China is the largest producer and consumer of tobacco in the world. Considering the constantly growing urban proportion, persuasive tobacco control measures are important in urban communities. Television, as one of the most pervasive mass media, can be used for this purpose. Methods The anti-smoking advertisement was carried out in five different time slots per day from 15 May to 15 June in 2011 across 12 channels of Chongqing TV. A cross-sectional study was conducted in the main municipal areas of Chongqing. A questionnaire was administered in late June to 1,342 native residents aged 18–45, who were selected via street intercept survey. Results Respondents who recognized the advertisement (32.77 %) were more likely to know or believe that smoking cigarettes caused impotence than those who did not recognize the advertisement (26.11 %). According to 25.5 % of smokers, the anti-smoking TV advertising made them consider quitting smoking. However, females (51.7 %) were less likely to be affected by the advertisement to stop and think about quitting smoking compared to males (65.6 %) (OR = 0.517, 95 % CI [0.281–0.950]). In addition, respondents aged 26–35 years (67.4 %) were more likely to try to persuade others to quit smoking than those aged 18–25 years (36.3 %) (OR = 0.457, 95 % CI [0.215–0.974]). Furthermore, non-smokers (87.4 %) were more likely to find the advertisement relevant than smokers (74.8 %) (OR = 2.34, 95 % CI [1.19–4.61]). Conclusions This study showed that this advertisement did not show significant differences on smoking-related knowledge and attitude between non-smokers who had seen the ad and those who had not. Thus, this form may not be the right tool to facilitate change in non-smokers. The ad should instead be focused on the smoking population. Gender, smoking status, and age influenced the effect of anti-smoking TV advertising on the general population in China.