Mitochondrial complex II, or succinate dehydrogenase (SDH), is regarded as a central regulator of respiratory adaptation and metabolic reprogramming in various stimuli and abnormalities. Four subunits of complex II are considered as tumor suppressors, whose mutations are associated with various type of cancer. However, little is known how complex II regulates cell proliferation. 2-Thenoyltrifluoroacetone (TTFA), an inhibitor of mitochondrial complex II, and SDHB shRNA were used to abolish the activity of complex II in cell lines. Inhibition the activity of complex II by TTFA treatment or knockdown of SDHB could trigger mitochondrial fragmentation and subsequently mitophagy. We also found that inhibition of complex II also increased the glucose consumption and the lactate production which termed as Warburg effect. Despite of these, complex II dysfunction showed negative regulation to cell proliferation. Collectively, complex II is a potential target to induce mitophagy and inhibit cell proliferation.