In the U.S. state of Kentucky, U.S. Route 60 is concurrent with US 51 and US 62 from the Ohio River bridge to the town of Wickliffe. At Wickliffe, US 60 separates from the other routes and heads generally northeast toward the city of Paducah. Between Wickliffe and Paducah, the towns of Barlow, La Center and Kevil are situated along the route.

In Paducah, US 60 intersects with Interstate 24, and I-24's business loop enters the city concurrent with US 60. In the downtown area, the route once again meets US 62, and also meets US 45. Once out of the city of Paducah, US 60 again veers to the northeast, generally following the Ohio River until reaching Smithland, where the route again turns to the east, passing through the small town of Burna. From Burna, U.S. 60 passes through Salem.

The city along the route is Marion. At Marion, US 60 turns once more to the north, where it heads toward Sturgis. From Sturgis, the route continues generally northward to Morganfield. In recent years, a by-pass of US 60 around the south and east sides of Morganfield has taken a great deal of traffic congestion out of the city.

US 60 passes through Waverly and Corydon before reaching Henderson. At Henderson, the route intersects Kentucky Route 136, Kentucky Route 425 and U.S. Route 41 Alternate. US 41-A is concurrent with US 60 along Green Street in the city of Henderson as it intersects with Kentucky Route 812 and Kentucky Route 351. At the U.S. Route 41/Pennyrile Parkway interchange, US 41-A ends and US 60 continues alone, by-passing the cities of Owensboro, Lewisport, Hawesville, and Hardinsburg.

In Fort Knox, US 60 connects with US 31W, and they remain together until downtown Louisville. Originally built as a "bypass route" around downtown Louisville, Alt US 60 used several existing roads running through Louisville to get between the east and south sides of town without having to travel through the heavily congested downtown or west ends of town. 'Alt 60' runs northeast to southwest from St. Matthews to Shively; including a stretch on one of Frederick Law Olmsted's last remaining parkways, Eastern Parkway. Before its completion and designation as Interstate 264 in 1984, the Henry Watterson Expressway connecting Dixie Highway (US 31W/US 60) and Shelbyville Rd (US 60) was signed Bypass U.S. 60.

After passing Shelbyville, it continues through the state capital of Frankfort, becomes four lanes and then heads more south than east toward Versailles. The more direct route to Lexington is two-lane US 421, Leestown Pike (once designated KY 50), which is technically southbound but runs more east than south in this area. In Versailles, US 60's eastbound lanes are very briefly the westbound lanes of US 62, which runs more south than west at that point. After turning east and intersecting the Bluegrass Parkway, US 60 is one of the major routes through Lexington. It intersects I-75 and from there roughly parallels I-64 and goes to Winchester and passes very near the Mountain Parkway but doesn't intersect with it. It proceeds to Mount Sterling, Morehead, past Carter Caves State Resort Park, on to Grayson and finally to Ashland. US 23 South follows US 60 East into Catlettsburg, where they split at 35th St. US 23 South continues straight as a 4-lane highway, while US 60 East turns left on 35th St., passes Oakland Avenue, and enters Kenova, West Virginia via the Billy C. Clark Bridge.

Until 2010, US 60 passed through the city of Owensboro, while the US 60 By-Pass (Wendell H. Ford Expressway) skirted the city to its south. The city of Owensboro and state of Kentucky requested the decommissioning of 60 within the city, and AASHTO approved the request, making the expressway the main line of US 60. As of April 2011, signage has been changed, and US 60 now solely follows the expressway around Owensboro.

Especially in the eastern and central part of the state, US 60 has been largely replaced by Interstate 64 for long distance travel, since both routes follow each other through much of this area. However, several cites in this area rely on US 60 to connect them to the interstate. By contrast, in the western part of the state, US 60 is not paired with an interstate highway and serves a much more independent purpose, connecting communities located along and near the Ohio River.