The ad's narrator asks in Spanish "Who supports Barack Obama" before highlighting endorsements of the president from Chávez and Castro's niece, Mariela.
The Romney campaign had previously attacked the president after the State Department approved a travel visa for Mariela Castro, calling the decision a "slap in the face" of Cubans. The Obama campaign later dismissed Romney's criticism as "bluster."
The Romney ad goes on to highlight an internal email exchange at the Environmental Protection Agency (EPA).
"And to top it off, Obama's Environmental Protection Agency sent emails for Hispanic Heritage month with Che Guevara's photo," the narrator says.
The EPA later said the email was sent mistakenly by a staffer and apologized for its distribution, although Republicans pounced on the issue.
"Che was a cold-blooded murderer who killed so many innocent victims," Rep. Mario Diaz-Balart (R-Fla.) told Fox News. "This Administration has granted visas to Mariela Castro and other top Cuban regime officials, offered the Castro brothers unilateral concessions, and now sends an E-mail with a image of an anti-American mass murderer."
According to CNN, which first reported the commercial, the ad began airing earlier this week in Miami, where Republicans see the sizable Cuban population as a key to securing the state.
In a statement Thursday, the Obama campaign called the commercial "another sign of desperation."
“Mitt Romney continues to play Hugo Chavez’s game, giving Chavez the attention he thrives on and that he doesn’t deserve," said Obama spokesman Dan Restrepo. "Keeping America safe and advancing U.S. interests requires the kind of leadership President Obama has provided, not Romney’s bluster."