The incident occurred between 3:35pm and 3:53pm on Saturday March 9 on the Sheffield to Chapeltown service.
It is reported that a group of three men began fighting with a man once the train had left Meadowhall station.
A number of punches were thrown however no injuries have been reported.
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The three men are described as white, with one aged in his 40s and wearing a blue scarf.
The second is said to have been in his 30s and was wearing a red baseball cap and the third is described as bald and wearing glasses.
Officers are keen to speak to anyone who witnessed the fight and are urging them to get in touch.
Anyone with information can call BTP 0800 40 50 40 or text 61016 quoting reference number 375 of March 9.
Alternatively you can call Crimestoppers anonymously on 0800 555 111.