President Xi Jinping oversaw a memorial service with columns of black-clad mourners wearing a white flower on their jackets, braving freezing temperatures at a monument featuring statues of victims in the eastern city.
Sirens blared at the start of the ceremony, while a giant "peace bell" tolled as doves were released into the air.
According to China some 300,000 civilians and soldiers were killed in a frenzy of murder, torture, rape, arson and looting in the six weeks after the invading Japanese military entered Nanjing, then the capital city, on December 13, 1937.
Many in China say this symbolises Japan's unwillingness to completely atone for its wartime aggression.
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Officially, Japan concedes that "the killing of a large number of noncombatants, looting and other acts occurred" but says it is "difficult" to determine precise figures.
The issue receded during the Cold War but has re-emerged as China strikes an increasingly muscular stance under Xi, while critics say Japanese revisionists have grown bolder under conservative leader Shinzo Abe.
Liang Yunxiang, an international relations expert at Peking University, said Beijing wants to keep such memories alive as leverage against Japan in modern-day disputes such as maritime territorial squabbles.
"There are current conflicts between the two countries, so historical issues are re-emerging. All history is contemporary," Liang said.
"Japan thinks these historical issues should have ended but China keeps hammering them as it becomes more powerful."
Commemorations centred on the sombre and poignant memorial hall and museum in Nanjing, with soldiers carrying a large wreath at the site as Xi stood sombrely.
Fewer than 100 people designated as massacre "survivors" remain alive, however, and both sides have repeatedly expressed a desire to look forward and avoiding rocking their huge trade relationship.
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