The star says she knows what it is like to face "thousands of allegations" and that she does not have a "lynch-mob mentality".
The star, who was the same age as Jackson and rose to fame alongside him during the 1980s, was asked about the Leaving Neverland documentary, which aired earlier this year, in an interview with British Vogue.
On camera in the two-part programme, alleged victims James Safechuck and Wade Robson claimed in graphic detail that they were sexually abused by Jackson when they were children.
Madonna, who is set to release her 14th album, Madame X, in June, told the magazine she had not watched it but said she knows what it is like to have allegations "hurled at me that are not true".
She said: "I don't have a lynch-mob mentality, so in my mind, people are innocent until proven guilty.
"I've had a thousand accusations hurled at me that are not true. So my attitude when people tell me things about people is, 'Can you prove it?'"
Asked what would constitute proof of Jackson's guilt, she replied: "I don't know, I haven't seen the film.
"But I guess it would be people recounting actual events - but then, of course, people sometimes lie.
"So I always say, 'What's the agenda? What do people want out of this? Are there people asking for money, is there some kind of extortion thing happening?'
"I would take all of those things into consideration."
Madonna, who took Jackson as her date to the Oscars in 1991, said of the HBO and Channel 4 documentary: "I guess I'll get around to seeing it. I haven't seen it yet."
Jackson, who died in 2009 aged 50, denied all allegations that he was involved in abusing underage boys and was acquitted of molestation charges in 2005.
The star's brothers Tito, Marlon and Jackie, and his nephew Taj, have dismissed the allegations against the late singer.
His estate has also criticised Leaving Neverland, describing it as a "tabloid character assassination" that the star had "endured in life and now in death".
In a statement, they described Mr Robson and Mr Safechuck as "two perjurers". Both had made sworn statements when Jackson was alive that he had not abused them.
In March, singer Barbra Streisand was criticised for saying the men were "thrilled" to spend time at his Neverland ranch.
She later posted an apology online.
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