The Giallorossi youngsters fell to a 6-1 defeat at Stadio Tre Fontane.
Alessio Riccardi scored the only goal of the game for the home side, who were 3-0 down at half-time. Real Madrid scored twice in injury time to give the scoreline a harsh look.
Qualification for the knockout stages of the competition remains possible for Roma, after CSKA Moscow and Viktoria Plzen drew elsewhere on the day.
A victory in the Czech Republic on the final matchday will guarantee the side's place in the next phase of the competition.
See photos from the game in the gallery below.
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