Cell chirality is an intrinsic property shown as biased cell rotation or orientation. While the righthanded helix of actin is known essential, it is intriguing of how single form of molecular handedness can be manifested into diverse and even mirrored forms of cell chirality. Here, we found that cell nucleus rotates with clockwise (CW) bias with smaller projected area but reverses to anticlockwise (ACW) bias when cell spreading increases. Actin analysis suggests that polymerization of righthanded radial fibers accounts for the ACW bias, while the CW bias is driven by the retrograde flow of CW-swirling transverse arc, which originates from tethered myosin II that simultaneously connect radial fibers and transverse arc. Thus, by tilting the balance between these two classes of actin fibers via cell spreading area or the main factors of actin, i.e., myosin II, α-actinin-1, Tpm4, and mDia2, the single form of actin helix can be unfolded into cell chirality with opposite bias. This finding suggests a mechanistic insight of cell chirality reversal, providing new perspective in mechanobiology and tissue formation.